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  • ADP-Ribosylation Levels and Patterns Correlate with Gene Expression and Clinical Outcomes in Ovarian Cancers
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Lesley B. Conrad, Ken Y. Lin, Tulip Nandu, Bryan A. Gibson, Jayanthi S. Lea, W. Lee Kraus

    Inhibitors of nuclear PARP enzymes (e.g., PARP-1) have improved clinical outcomes in ovarian cancer, especially in patients with BRCA1/2 gene mutations or additional homologous recombination (HR) DNA repair pathway deficiencies. These defects serve as biomarkers for response to PARP inhibitors (PARPi). We sought to identify an additional biomarker that could predict responses to both conventional chemotherapy and PARPi in ovarian cancers. We focused on cellular ADP-ribosylation (ADPRylation), which is catalyzed by PARP enzymes and detected by detection reagents we developed previously. We determined molecular phenotypes of 34 high-grade serous ovarian cancers and associated them with clinical outcomes. We used the levels and patterns of ADPRylation and PARP-1 to distribute ovarian cancers into distinct molecular phenotypes, which exhibit dramatically different gene expression profiles. In addition, the levels and patterns of ADPRylation, PARP-1 protein, and gene expression correlated with clinical outcomes in response to platinum-based chemotherapy, with cancers exhibiting the highest levels of ADPRylation having the best outcomes independent of BRCA1/2 status. Finally, in cell culture-based assays using patient-derived ovarian cancer cell lines, ADPRylation levels correlated with sensitivity to the PARPi, Olaparib, with cell lines exhibiting high levels of ADPRylation having greater sensitivity to Olaparib. Collectively, our study demonstrates that ovarian cancers exhibit a wide range of ADPRylation levels, which correlate with therapeutic responses and clinical outcomes. These results suggest ADPRylation may be a useful biomarker for PARPi sensitivity in ovarian cancers, independent of BRCA1/2 or homologous recombination deficiency status.

    更新日期:2020-01-16
  • Predictive and Pharmacodynamic Biomarkers of Response to the Phosphatidylinositol 3-Kinase Inhibitor Taselisib in Breast Cancer Preclinical Models
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Heather M. Moore, Heidi M. Savage, Carol O'Brien, Wei Zhou, Ethan S. Sokol, Michael E. Goldberg, Ciara Metcalfe, Lori S. Friedman, Mark R. Lackner, Timothy R. Wilson

    The PI3K signaling pathway serves as a central node in regulating cell survival, proliferation, and metabolism. PIK3CA , the gene encoding the PI3K catalytic subunit p110-alpha, is commonly altered in breast cancer resulting in the constitutive activation of the PI3K pathway. Using an unbiased cell line screening approach, we tested the sensitivity of breast cancer cell lines to taselisib, a potent PI3K inhibitor, and correlated sensitivity with key biomarkers ( PIK3CA , HER2, PTEN, and ESR1 ). We further assessed how taselisib modulates downstream signaling in the different genomic backgrounds that occur within breast cancer. We found that sensitivity to taselisib correlated with the presence of PIK3CA mutations, but was independent of HER2 status. We further showed that HER2-amplified/ PIK3CA wild-type cell lines are not as sensitive to taselisib when compared with HER2-amplified/ PIK3CA -mutant cell lines. In a PIK3CA- mutant/PTEN null background, PI3K downstream signaling rebounded in the presence of taselisib correlating with decreased sensitivity at later time points. Finally, we observed that PIK3CA mutations cooccurred with mutations in the estrogen receptor (ER; ESR1 ) in metastatic tumors from patients with ER+ breast cancer. However, the cooccurrence of an ESR1 mutation with a PIK3CA mutation did not affect response to taselisib in a single agent setting or in combination with fulvestrant. In summary, these data suggest that development of taselisib in breast cancer should occur in a PIK3CA -mutant setting with cotreatments determined by the specific subtypes under investigation. This article is featured in Highlights of This Issue, [p. 1][1] [1]: /lookup/volpage/19/1?iss=1

    更新日期:2020-01-16
  • Pralatrexate in Combination with Oxaliplatin in Advanced Esophagogastric Cancer: A Phase II Trial with Predictive Molecular Correlates
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Usha Malhotra, Sarbajit Mukherjee, Christos Fountzilas, Patrick Boland, Austin Miller, Santosh Patnaik, Kristopher Attwood, Sai Yendamuri, Araba Adjei, Eric Kannisto, Mateusz Opyrchal, Peter Bushunow, Peter Loud, Renuka Iyer, Nikhil Khushalani

    The purpose of our study was to evaluate the efficacy of a combination of pralatrexate plus oxaliplatin in advanced esophagogastric cancer (EGC), analyze the impact of polymorphisms in folate metabolism pathway genes on toxicity and efficacy of pralatrexate, and to evaluate microRNA profile of tumor epithelium as a predictive biomarker. This was a two-stage trial with a safety lead in cohort and a primary endpoint of overall response rate (ORR). Patients received biweekly intravenous oxaliplatin (85 mg/m2) and pralatrexate (Dose level 1 [D1], 120 mg/m2; dose level-1 [D-1] 100 mg/m2). Single-nucleotide polymorphisms (SNP) in genes encoding proteins involved in pralatrexate metabolism were evaluated in germline DNA. microRNA profiling of the tumor epithelium was performed. ORR was 26%. Dose-limiting toxicities were observed in 2 of 4 patients at D1 and none at D-1. The T>C polymorphism in DHFR rs11951910 was significantly associated with lower progression-free survival (PFS; P ≤ 0.01), whereas the presence of the SLC19A1 rs2838957 G>A polymorphism was associated with improved PFS ( P = 0.02). Presence of the GGH rs3780130 A>T and SLC19A1 rs1051266 G>A polymorphisms were significantly associated with better overall survival (OS; P = 0.01), whereas GGH rs7010484 T>C polymorphism was associated significantly with reduced OS ( P = 0.04). There was no correlation between epithelial microRNA expression profile with disease progression or response. We conclude that the combination of oxaliplatin and pralatrexate is safe, is well tolerated, and has modest efficacy in advanced EGC. Pharmacogenomic analysis may be relevant to the use of pralatrexate in combination with platinum agents.

    更新日期:2020-01-16
  • Identification of Expression Profiles Defining Distinct Prognostic Subsets of Radioactive-Iodine Refractory Differentiated Thyroid Cancer from the DECISION Trial
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Jaume Capdevila, Ignacio Matos, Francesco M. Mancuso, Carmela Iglesias, Paolo Nuciforo, Carles Zafon, Hector G. Palmer, Zighereda Ogbah, Laura Muiños, Jorge Hernando, Guillermo Villacampa, Carol E. Peña, Josep Tabernero, Marcia S. Brose, Martin Schlumberger, Ana Vivancos

    Several biomarkers have been suggested to have prognostic value in differentiated thyroid carcinomas (DTC) with no validation in the refractory setting, including all tumor subtypes. We aim to correlate RNA expression profiles with survival based on patients included in the DECISION trial. We obtained 247 samples from the 417 patients included in the DECISION study and performed RNAseq analysis (77 million paired-end reads for each sample on HiSeq2000). After quality control, 125 samples were included in the secondary analysis and mapped against the human reference genome (GRCh38) with STAR (v2.5.1b) using ENCODE parameter. Survival analysis was calculated using the Kaplan–Meier method and log-rank test was used for statistical comparison. In this post hoc analysis, we identified three groups of tumors based on their gene expression profile: BRAF-like, RAS-like, and non-BRAF-non-RAS-like (NoBRaL). No significant correlation with sorafenib responders was observed. However, we identified a statistically significant correlation between the RNA-expression profiles and progression-free survival. The BRAF-like profile had a significantly better outcome compared with RAS-like and NoBRaL (11.8, 6.2, and 5.5 months, respectively) [HR: 0.31, 95% confidence interval (CI), 0.17–0.60; P < 0.001 and HR: 0.36 (95% CI, 0.21–0.63); P < 0.001] and HR: 0.36 (95% CI, 0.21–0.63; P < 0.001) and maintained significance as an independent prognostic factor for overall survival in the multivariate analysis for papillary thyroid cancers. To our knowledge, this is the first comprehensive RNA-seq analysis of all histologic subtypes of DTC. The RNA expression profiles identified may suggest a new prognostic parameter to be considered before recommendation of systemic therapies or the design of stratification factors for future clinical trials.

    更新日期:2020-01-16
  • Extracellular vesicle-mediated in vitro transcribed mRNA delivery for treatment of HER2+ breast cancer xenografts in mice by prodrug CB1954 without general toxicity
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-15
    Alexis V. Forterre, Jing-Hung Wang, Alain Delcayre, Kyuri Kim, Carol Green, Mark D Pegram, Stefanie S Jeffrey, A.C. Matin

    Prodrugs are harmless until activated by a bacterial or viral gene product; they constitute the basis of gene delivered prodrug therapies called GDEPTs, which can kill tumors without major side effects. Previously, we utilized the prodrug CNOB (not clinically tested) and enzyme HChrR6 in GDEPT to generate the drug MCHB in tumors. Extracellular vesicles (EVs) were used for directed gene delivery and HChrR6 mRNA as gene. Here, the clinical transfer of this approach is enhanced by: 1. Use of CB1954 (tretazicar) for which safe human dose is established; HChrR6 can activate this prodrug. 2. EVs delivered in vitro transcribed (IVT) HChrR6 mRNA, eliminating the potentially harmful plasmid transfection of EV-producer cells we utilized previously; this has not been done before. IVT mRNA loading of EVs required several steps. Naked mRNA being unstable, we ensured its prodrug activating functionality at each step. This was not possible using tretazicar itself; we relied instead on HChrR6's ability to convert CNOB into MCHB, whose fluorescence is easily visualizable. HChrR6 mRNA-translated product's ability to generate fluorescence from CNOB vicariously indicated its competence for tretazicar activation. 3. Systemic IVT mRNA loaded EVs displaying an anti-HER2 scFv ("IVT EXO-DEPTs") and tretazicar caused growth arrest of human HER2+ breast cancer xenografts in athymic mice. As this occurred without injury to other tissues, absence of off-target mRNA delivery is strongly indicated. Many cancer sites are not amenable for direct gene injection, but current GDEPTs require this. In circumventing this need, a major advance in GDEPT applicability has been accomplished.

    更新日期:2020-01-16
  • Preclinical activity of ribociclib in squamous cell carcinoma of the head and neck
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-10
    Gabrielle van Caloen, Sandra Schmitz, Mariama El Baroudi, Xavier Caignet, Sébastien Pyr dit Ruys, Pierre P Roger, Didier Vertommen, Jean-Pascal Machiels

    Purpose: Cell cycle pathway impairments resulting in cyclin-dependent kinase (CDK) 4 and 6 activation are frequently observed in human papillomavirus (HPV)-negative squamous cell carcinoma of the head and neck (SCCHN). We investigated the activity of ribociclib, a CDK4/6 inhibitor, in SCCHN models with the aim of identifying predictive biomarkers of response. Methods: HPV-negative or HPV-positive SCCHN cell lines (n=8) and patient-derived tumor xenograft (PDTX) models (n=6) were used. The models were classified according to their sensitivity to ribociclib to investigate potential predictive biomarkers. Results: Ribociclib had a cytostatic effect in some HPV-negative SCCHN models but had no effect in HPV-positive models. In SCCHN cell lines and PDTXs, the retinoblastoma (Rb) protein expression level correlated with ribociclib activity. Rb knockdown was, however, not sufficient to block G0-G1 arrest induced by ribociclib in Detroit-562 where p107, p130 and Forkhead BOX M1 (FOXM1) were also implicated in ribociclib activity. Cell lines harboring epithelial-mesenchymal transition (EMT) features were less sensitive to ribociclib than those with an epithelial phenotype. Rb downregulation induced EMT in our Rb-expressing SCCHN cell lines. However, ribociclib still had significant activity in one PDTX model with high Rb and vimentin expression, suggesting that the presence of vimentin alone is not enough to induce ribociclib resistance. Conclusion: These findings suggest that CDK4/6 inhibitors should be investigated in patients with HPV-negative SCCHN with high Rb expression and an epithelial phenotype. Although these biomarkers are not predictive in all cases, they may enrich the population that could benefit from CDK4/6 inhibitors.

    更新日期:2020-01-10
  • KLF4 p.A472D Mutation Contributes to Acquired Resistance to Cetuximab in Colorectal Cancer
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-10
    Song Ye, Xiaoge Hu, Chao Ni, Weiwei Jin, Yaping Xu, Lianpeng Chang, Huaixiang zhou, Jiahong Jiang, Liu Yang

    With the increase of treatment course, resistance to epidermal growth factor receptor (EGFR) blockade is inevitable in patients with metastatic colorectal cancer (mCRC). KRAS mutations have been considered to be primary drivers of this resistance; however, the potential function of other genes has not been extensively investigated. This study collected 17 plasma samples from mCRC patients with cetuximab resistance, and target-capture deep sequencing was used to identify mutations in circulating tumor DNA (ctDNA). Analysis of mutational prevalence in ctDNA was performed from three CRC tissue-based datasets and one ctDNA dataset. The prevalence of mutations identified in ctDNA was consistent with both CRC tissue-based and ctDNA datasets. Clonal analysis revealed that 41.2% of patients were positive for at least one subclone. Multiple mechanisms of cetuximab resistance were co-existed in individual patients, and one of the patients even harbored nine distinct mutations. In particular, functional study of Krüppel-like factor 4 (KLF4) p.A472D revealed increased cetuximab resistance in CRC cells, which was associated with the increased phosphorylation of downstream EGFR signaling proteins. These results suggest that the KLF4 p.A472D may contribute to cetuximab resistance in patients with mCRC and thus, it may serve as a new biomarker in clinical application. Monitoring somatic mutations related to cetuximab resistance in mCRC patients through ctDNA may provide real-time insights for clinical reference and treatment planning.

    更新日期:2020-01-10
  • Efficacy of FGFR inhibitors and combination therapies for acquired resistance in FGFR2-fusion cholangiocarcinoma
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-07
    Melanie A. Krook, Alexandria Lenyo, Max Wilberding, Hannah Barker, Mikayla Dantuono, Kelly M Bailey, Hui-Zi Chen, Julie W Reeser, Michele R. Wing, Jharna Miya, Eric Samorodnitsky, Amy M. Smith, Thuy Dao, Dorrelyn M Martin, Kristen K Ciombor, John Hays, Aharon G Freud, Sameek Roychowdhury

    The fibroblast growth factor receptor (FGFR) signaling pathway is aberrantly activated in approximately 15-20% of intrahepatic cholangiocarcinoma patients. Currently, several FGFR kinase inhibitors are being assessed in clinical trials for patients with FGFR-altered cholangiocarcinoma. Despite evidence of initial responses and disease control, virtually all patients eventually develop acquired resistance. Thus, there is a critical need for the development of innovative therapeutic strategies to overcome acquired drug resistance. Here, we present findings from a patient with FGFR2-altered metastatic cholangiocarcinoma who enrolled in a phase II clinical trial of the FGFR inhibitor, infigratinib (BGJ398). Treatment was initially effective as demonstrated by imaging and tumor marker response; however, after eight months on trial, the patient exhibited tumor regrowth and disease progression. Targeted sequencing of tumor DNA after disease progression revealed the FGFR2 kinase domain p.E565A and p.L617M single nucleotide variants (SNVs) hypothesized to drive acquired resistance to infigratinib. The sensitivities of these FGFR2 SNVs, which were detected post-infigratinib therapy, were extended to include clinically relevant FGFR inhibitors including AZD4547, erdafitinib (JNJ-42756493), dovitinib, ponatinib, and TAS120, and were evaluated in vitro. Through a proteomics approach, we identified upregulation of the PI3K/AKT/mTOR signaling pathway in cells harboring the FGFR2 p.E565A mutation and demonstrated that combination therapy strategies with FGFR and mTOR inhibitors may be used to overcome resistance to FGFR inhibition, specific to infigratinib. Collectively, these studies support the development of novel combination therapeutic strategies in addition to the next generation of FGFR inhibitors to overcome acquired resistance in patients.

    更新日期:2020-01-08
  • 10D1F, an Anti-HER3 Antibody that Uniquely Blocks the Receptor Heterodimerization Interface, Potently Inhibits Tumor Growth Across a Broad Panel of Tumor Models
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-07
    Dipti Thakkar, Vicente Sancenon, Marvin M. Taguiam, Siyu Guan, Zhihao Wu, Eric Ng, Konrad H. Paszkiewicz, Piers J. Ingram, Jerome D. Boyd-Kirkup

    In recent years, HER3 has increasingly been implicated in the progression of a variety of tumor types and in acquired resistance to EGFR and HER2 therapies. Whereas EGFR and HER2 primarily signal through the MAPK pathway, HER3, as a heterodimer with EGFR or HER2, potently activates the PI3K pathway. Despite its critical role, previous attempts to target HER3 with neutralizing antibodies have shown disappointing efficacy in the clinic, most likely due to suboptimal and indirect mechanisms of action that fail to completely block heterodimerization; for example, tumors can escape inhibition of ligand binding by upregulating ligand-independent mechanisms of HER3 activation. We therefore developed 10D1F, a picomolar affinity, highly specific anti-HER3 neutralizing antibody that binds the HER3 heterodimerization interface, a region that was hitherto challenging to raise antibodies against. We demonstrate that 10D1F potently inhibits both EGFR:HER3 and HER2:HER3 heterodimerization to durably suppress activation of the PI3K pathway in a broad panel of tumor models. Even as a monotherapy, 10D1F shows superior inhibition of tumor growth in the same cell lines both in vitro and in mouse xenograft experiments, when compared with other classes of anti-HER3 antibodies. This includes models demonstrating ligand-independent activation of heterodimerization as well as constitutively activating mutations in the MAPK pathway. Possessing favorable pharmacokinetic and toxicologic profiles, 10D1F uniquely represents a new class of anti-HER3 neutralizing antibodies with a novel mechanism of action that offers significant potential for broad clinical benefit. 10D1F is a novel anti-HER3 antibody that uniquely binds the receptor dimerization interface to block ligand-dependent and independent heterodimerization with EGFR/HER2 and thus more potently inhibits tumor growth than existing anti-HER3 antibodies.

    更新日期:2020-01-08
  • Selected Articles from This Issue
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    American Association for Cancer Research

    ### [Hingorani et al. Page 157][1] Although cytotoxic chemotherapies improve the control of locally advanced, non-metastatic tumors with radiotherapy, their lack of targeting can increase the damage done to nearby normal tissue and induce systemic toxicities. To achieve a more targeted

    更新日期:2020-01-04
  • The Ongoing Search for Biomarkers of CDK4/6 Inhibitor Responsiveness in Breast Cancer
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Scott F. Schoninger, Stacy W. Blain

    CDK4 inhibitors (CDK4/6i), such as palbociclib, ribociclib, and abemaciclib, are approved in combination with hormonal therapy as a front-line treatment for metastatic HR+, HER2- breast cancer. Their targets, CDK4 and CDK6, are cell-cycle regulatory proteins governing the G1–S phase transition across many tissue types. A key challenge remains to uncover biomarkers to identify those patients that may benefit from this class of drugs. Although CDK4/6i addition to estrogen modulation therapy essentially doubles the median progression-free survival, overall survival is not significantly increased. However, in reality only a subset of treated patients respond. Many patients exhibit primary resistance to CDK4/6 inhibition and do not derive any benefit from these agents, often switching to chemotherapy within 6 months. Some patients initially benefit from treatment, but later develop secondary resistance. This highlights the need for complementary or companion diagnostics to pinpoint patients who would respond. In addition, because CDK4 is a bona fide target in other tumor types where CDK4/6i therapy is currently in clinical trials, the lack of target identification may obscure benefit to a subset of patients there as well. This review summarizes the current status of CDK4/6i biomarker test development, both in clinical trials and at the bench, with particular attention paid to those which have a strong biological basis as well as supportive clinical data. This article is featured in Highlights of This Issue, [p. 1][1] [1]: /lookup/volpage/19/1?iss=1

    更新日期:2020-01-04
  • Pharmacology of the ATM Inhibitor AZD0156: Potentiation of Irradiation and Olaparib Responses Preclinically
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Lucy C. Riches, Antonio G. Trinidad, Gareth Hughes, Gemma N. Jones, Adina M. Hughes, Andrew G. Thomason, Paul Gavine, Andy Cui, Stephanie Ling, Jonathan Stott, Roger Clark, Samantha Peel, Pendeep Gill, Louise M. Goodwin, Aaron Smith, Kurt G. Pike, Bernard Barlaam, Martin Pass, Mark J. O'Connor, Graeme Smith, Elaine B. Cadogan

    AZD0156 is a potent and selective, bioavailable inhibitor of ataxia-telangiectasia mutated (ATM) protein, a signaling kinase involved in the DNA damage response. We present preclinical data demonstrating abrogation of irradiation-induced ATM signaling by low doses of AZD0156, as measured by phosphorylation of ATM substrates. AZD0156 is a strong radiosensitizer in vitro , and using a lung xenograft model, we show that systemic delivery of AZD0156 enhances the tumor growth inhibitory effects of radiation treatment in vivo . Because ATM deficiency contributes to PARP inhibitor sensitivity, preclinically, we evaluated the effect of combining AZD0156 with the PARP inhibitor olaparib. Using ATM isogenic FaDu cells, we demonstrate that AZD0156 impedes the repair of olaparib-induced DNA damage, resulting in elevated DNA double-strand break signaling, cell-cycle arrest, and apoptosis. Preclinically, AZD0156 potentiated the effects of olaparib across a panel of lung, gastric, and breast cancer cell lines in vitro , and improved the efficacy of olaparib in two patient-derived triple-negative breast cancer xenograft models. AZD0156 is currently being evaluated in phase I studies ([NCT02588105][1]). [1]: /lookup/external-ref?link_type=CLINTRIALGOV&access_num=NCT02588105&atom=%2Fmolcanther%2F19%2F1%2F13.atom

    更新日期:2020-01-04
  • The Novel ATR Inhibitor BAY 1895344 Is Efficacious as Monotherapy and Combined with DNA Damage-Inducing or Repair-Compromising Therapies in Preclinical Cancer Models
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Antje M. Wengner, Gerhard Siemeister, Ulrich Lücking, Julien Lefranc, Lars Wortmann, Philip Lienau, Benjamin Bader, Ulf Bömer, Dieter Moosmayer, Uwe Eberspächer, Sven Golfier, Christoph A. Schatz, Simon J. Baumgart, Bernard Haendler, Pascale Lejeune, Andreas Schlicker, Franz von Nussbaum, Michael Brands, Karl Ziegelbauer, Dominik Mumberg

    The DNA damage response (DDR) secures the integrity of the genome of eukaryotic cells. DDR deficiencies can promote tumorigenesis but concurrently may increase dependence on alternative repair pathways. The ataxia telangiectasia and Rad3-related (ATR) kinase plays a central role in the DDR by activating essential signaling pathways of DNA damage repair. Here, we studied the effect of the novel selective ATR kinase inhibitor BAY 1895344 on tumor cell growth and viability. Potent antiproliferative activity was demonstrated in a broad spectrum of human tumor cell lines. BAY 1895344 exhibited strong monotherapy efficacy in cancer xenograft models that carry DNA damage repair deficiencies. The combination of BAY 1895344 with DNA damage–inducing chemotherapy or external beam radiotherapy (EBRT) showed synergistic antitumor activity. Combination treatment with BAY 1895344 and DDR inhibitors achieved strong synergistic antiproliferative activity in vitro , and combined inhibition of ATR and PARP signaling using olaparib demonstrated synergistic antitumor activity in vivo . Furthermore, the combination of BAY 1895344 with the novel, nonsteroidal androgen receptor antagonist darolutamide resulted in significantly improved antitumor efficacy compared with respective single-agent treatments in hormone-dependent prostate cancer, and addition of EBRT resulted in even further enhanced antitumor efficacy. Thus, the ATR inhibitor BAY 1895344 may provide new therapeutic options for the treatment of cancers with certain DDR deficiencies in monotherapy and in combination with DNA damage–inducing or DNA repair–compromising cancer therapies by improving their efficacy.

    更新日期:2020-01-04
  • A Multifunctional Therapy Approach for Cancer: Targeting Raf1- Mediated Inhibition of Cell Motility, Growth, and Interaction with the Microenvironment
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Limin Zhang, Abhinandan Pattanayak, Wenqi Li, Hyun-Kyung Ko, Graham Fowler, Ryan Gordon, Raymond Bergan

    Prostate cancer cells move from their primary site of origin, interact with a distant microenvironment, grow, and thereby cause death. It had heretofore not been possible to selectively inhibit cancer cell motility. Our group has recently shown that inhibition of intracellular activation of Raf1 with the small-molecule therapeutic KBU2046 permits, for the first time, selective inhibition of cell motility. We hypothesized that simultaneous disruption of multiple distinct functions that drive progression of prostate cancer to induce death would result in advanced disease control. Using a murine orthotopic implantation model of human prostate cancer metastasis, we demonstrate that combined treatment with KBU2046 and docetaxel retains docetaxel's antitumor action, but provides improved inhibition of metastasis, compared with monotherapy. KBU2046 does not interfere with hormone therapy, inclusive of enzalutamide-mediated inhibition of androgen receptor (AR) function and cell growth inhibition, and inclusive of the ability of castration to inhibit LNCaP-AR cell outgrowth in mice. Cell movement is necessary for osteoclast-mediated bone degradation. KBU2046 inhibits Raf1 and its downstream activation of MEK1/2 and ERK1/2 in osteoclasts, inhibiting cytoskeleton rearrangement, resorptive cavity formation, and bone destruction in vitro , with improved effects observed when the bone microenvironment is chemically modified by pretreatment with zoledronic acid. Using a murine cardiac injection model of human prostate cancer bone destruction quantified by CT, KBU2046 plus zoledronic exhibit improved inhibitory efficacy, compared with monotherapy. The combined disruption of pathways that drive cell movement, interaction with bone, and growth constitutes a multifunctional targeting strategy that provides advanced disease control. This article is featured in Highlights of This Issue, [p. 1][1] [1]: /lookup/volpage/19/1?iss=1

    更新日期:2020-01-04
  • Quantitative High-Throughput Screening Using an Organotypic Model Identifies Compounds that Inhibit Ovarian Cancer Metastasis
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Hilary A. Kenny, Madhu Lal-Nag, Min Shen, Betul Kara, Dominik A. Nahotko, Kristen Wroblewski, Sarah Fazal, Siquan Chen, Chun-Yi Chiang, Yen-Ju Chen, Kyle R. Brimacombe, Juan Marugan, Marc Ferrer, Ernst Lengyel

    The tumor microenvironment (TME) is a key determinant of metastatic efficiency. We performed a quantitative high-throughput screen (qHTS) of diverse medicinal chemistry tractable scaffolds (44,420 compounds) and pharmacologically active small molecules (386 compounds) using a layered organotypic, robust assay representing the ovarian cancer metastatic TME. This 3D model contains primary human mesothelial cells, fibroblasts, and extracellular matrix, to which fluorescently labeled ovarian cancer cells are added. Initially, 100 compounds inhibiting ovarian cancer adhesion/invasion to the 3D model in a dose-dependent manner were identified. Of those, eight compounds were confirmed active in five high-grade serous ovarian cancer cell lines and were further validated in secondary in vitro and in vivo biological assays. Two tyrosine kinase inhibitors, PP-121 and milciclib, and a previously unreported compound, NCGC00117362, were selected because they had potency at 1 μmol/L in vitro . Specifically, NCGC00117362 and PP-121 inhibited ovarian cancer adhesion, invasion, and proliferation, whereas milciclib inhibited ovarian cancer invasion and proliferation. Using in situ kinase profiling and immunoblotting, we found that milciclib targeted Cdk2 and Cdk6, and PP-121 targeted mTOR. In vivo , all three compounds prevented ovarian cancer adhesion/invasion and metastasis, prolonged survival, and reduced omental tumor growth in an intervention study. To evaluate the clinical potential of NCGC00117362, structure–activity relationship studies were performed. Four close analogues of NCGC00117362 efficiently inhibited cancer aggressiveness in vitro and metastasis in vivo . Collectively, these data show that a complex 3D culture of the TME is effective in qHTS. The three compounds identified have promise as therapeutics for prevention and treatment of ovarian cancer metastasis.

    更新日期:2020-01-04
  • Inhibition of Autotaxin with GLPG1690 Increases the Efficacy of Radiotherapy and Chemotherapy in a Mouse Model of Breast Cancer
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Xiaoyun Tang, Melinda Wuest, Matthew G.K. Benesch, Jennifer Dufour, YuanYuan Zhao, Jonathan M. Curtis, Alain Monjardet, Bertrand Heckmann, David Murray, Frank Wuest, David N. Brindley

    Autotaxin catalyzes the formation of lysophosphatidic acid, which stimulates tumor growth and metastasis and decreases the effectiveness of cancer therapies. In breast cancer, autotaxin is secreted mainly by breast adipocytes, especially when stimulated by inflammatory cytokines produced by tumors. In this work, we studied the effects of an ATX inhibitor, GLPG1690, which is in phase III clinical trials for idiopathic pulmonary fibrosis, on responses to radiotherapy and chemotherapy in a syngeneic orthotopic mouse model of breast cancer. Tumors were treated with fractionated external beam irradiation, which was optimized to decrease tumor weight by approximately 80%. Mice were also dosed twice daily with GLPG1690 or vehicle beginning at 1 day before the radiation until 4 days after radiation was completed. GLPG1690 combined with irradiation did not decrease tumor growth further compared with radiation alone. However, GLPG1690 decreased the uptake of 3′-deoxy-3′-[18F]-fluorothymidine by tumors and the percentage of Ki67-positive cells. This was also associated with increased cleaved caspase-3 and decreased Bcl-2 levels in these tumors. GLPG1690 decreased irradiation-induced C-C motif chemokine ligand-11 in tumors and levels of IL9, IL12p40, macrophage colony-stimulating factor, and IFNγ in adipose tissue adjacent to the tumor. In other experiments, mice were treated with doxorubicin every 2 days after the tumors developed. GLPG1690 acted synergistically with doxorubicin to decrease tumor growth and the percentage of Ki67-positive cells. GLPG1690 also increased 4-hydroxynonenal-protein adducts in these tumors. These results indicate that inhibiting ATX provides a promising adjuvant to improve the outcomes of radiotherapy and chemotherapy for breast cancer.

    更新日期:2020-01-04
  • A Novel Small Molecule Targets Androgen Receptor and Its Splice Variants in Castration-Resistant Prostate Cancer
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Zhenyu Yang, Dan Wang, James K. Johnson, Laura E. Pascal, Keita Takubo, Raghunandan Avula, Anish Bhaswanth Chakka, Jianhua Zhou, Wei Chen, Mingming Zhong, Qiong Song, Hui Ding, Zeyu Wu, Uma R. Chandran, Taber S. Maskrey, Joel B. Nelson, Peter Wipf, Zhou Wang

    Reactivation of androgen receptor (AR) appears to be the major mechanism driving the resistance of castration-resistant prostate cancer (CRPC) to second-generation antiandrogens and involves AR overexpression, AR mutation, and/or expression of AR splice variants lacking ligand-binding domain. There is a need for novel small molecules targeting AR, particularly those also targeting AR splice variants such as ARv7. A high-throughput/high-content screen was previously reported that led to the discovery of a novel lead compound, 2-(((3,5-dimethylisoxazol-4-yl)methyl)thio)-1-(4-(2,3-dimethylphenyl)piperazin-1-yl)ethan-1-one (IMTPPE), capable of inhibiting nuclear AR level and activity in CRPC cells, including those resistant to enzalutamide. A novel analogue of IMTPPE, JJ-450, has been investigated with evidence for its direct and specific inhibition of AR transcriptional activity via a pulldown assay and RNA-sequencing analysis, PSA-based luciferase, qPCR, and chromatin immunoprecipitation assays, and xenograft tumor model 22Rv1. JJ-450 blocks AR recruitment to androgen-responsive elements and suppresses AR target gene expression. JJ-450 also inhibits ARv7 transcriptional activity and its target gene expression. Importantly, JJ-450 suppresses the growth of CRPC tumor xenografts, including ARv7-expressing 22Rv1. Collectively, these findings suggest JJ-450 represents a new class of AR antagonists with therapeutic potential for CRPC, including those resistant to enzalutamide.

    更新日期:2020-01-04
  • Verticillin A Causes Apoptosis and Reduces Tumor Burden in High-Grade Serous Ovarian Cancer by Inducing DNA Damage
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Amrita Salvi, Chiraz Soumia M. Amrine, Julia R. Austin, KiAundra Kilpatrick, Angela Russo, Daniel Lantvit, Esther Calderon-Gierszal, Zachary Mattes, Cedric J. Pearce, Mark W. Grinstaff, Aaron H. Colby, Nicholas H. Oberlies, Joanna E. Burdette

    High-grade serous ovarian cancer (HGSOC) is the most lethal gynecological malignancy in women worldwide and the fifth most common cause of cancer-related deaths among U.S. women. New therapies are needed to treat HGSOC, particularly because most patients develop resistance to current first-line therapies. Many natural product and fungal metabolites exhibit anticancer activity and represent an untapped reservoir of potential new agents with unique mechanism(s) of action. Verticillin A, an epipolythiodioxopiperazine alkaloid, is one such compound, and our recent advances in fermentation and isolation are now enabling evaluation of its anticancer activity. Verticillin A demonstrated cytotoxicity in HGSOC cell lines in a dose-dependent manner with a low nmol/L IC50. Furthermore, treatment with verticillin A induced DNA damage and caused apoptosis in HGSOC cell lines OVCAR4 and OVCAR8. RNA-Seq analysis of verticillin A–treated OVCAR8 cells revealed an enrichment of transcripts in the apoptosis signaling and the oxidative stress response pathways. Mass spectrometry histone profiling confirmed reports that verticillin A caused epigenetic modifications with global changes in histone methylation and acetylation marks. To facilitate in vivo delivery of verticillin A and to monitor its ability to reduce HGSOC tumor burden, verticillin A was encapsulated into an expansile nanoparticle (verticillin A-eNP) delivery system. In an in vivo human ovarian cancer xenograft model, verticillin A-eNPs decreased tumor growth and exhibited reduced liver toxicity compared with verticillin A administered alone. This study confirmed that verticillin A has therapeutic potential for treatment of HGSOC and that encapsulation into expansile nanoparticles reduced liver toxicity.

    更新日期:2020-01-04
  • A Novel Salicylanilide Derivative Induces Autophagy Cell Death in Castration-Resistant Prostate Cancer via ER Stress-Activated PERK Signaling Pathway
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Chia-Ling Hsieh, Hsu-Shan Huang, Kuan-Chou Chen, Teigi Saka, Chih-Ying Chiang, Leland W.K. Chung, Shian-Ying Sung

    Metastatic castration-resistant prostate cancer (CRPC) is currently incurable. Cancer growth and progression is intimately affected by its interaction with host microenvironment. Cotargeting of the stroma and prostate cancer is therefore an emerging therapeutic strategy for metastatic CRPC. Cancer-induced osteoclastogenesis is known to contribute to CRPC bone metastasis. This study is to extend pharmacologic value of our synthesized LCC03, a derivative of 5-(2′,4′-difluorophenyl)-salicylanilide that has previously testified for its osteoclastogenesis activity, by exploring its additional cytotoxic properties and underlying mechanism in CRPC cells. LCC03 was chemically synthesized and examined for cell growth inhibition in a serial of CRPC cell lines. We demonstrated that LCC03 dose-dependently suppressed proliferation and retarded cell-cycle progression in CRPC cells. The classical autophagy features, including autophagosome formation and LC3-II conversion, were dramatically shown in LCC03-treated CRPC cells, and it was associated with the suppressed AKT/mTOR signaling pathways, a major negative regulator of autophagy. Moreover, an expanded morphology of the endoplasmic reticulum (ER), increased expression of the ER stress markers GRP78 and PERK, and eIF2α phosphorylation were observed. Blockage of autophagy and PERK pathways using small molecule inhibitors or shRNA knockdown reversed LCC03-induced autophagy and cell death, thus indicating that the PERK–eIF2α pathway contributed to the LCC03-induced autophagy. Furthermore, treatment of tumor-bearing mice with intraperitoneal administered LCC03 suppressed the growth of CRPC xenografts in mouse bone without systemic toxicity. The dual action of 5-(2′,4′-difluorophenyl)-salicylanilide on targeting both the osteoclasts and the tumor cells strongly indicates that LCC03 is a promising anticancer candidate for preventing and treating metastatic CRPC.

    更新日期:2020-01-04
  • Molecular Inhibitor of QSOX1 Suppresses Tumor Growth In Vivo
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Amber L. Fifield, Paul D. Hanavan, Douglas O. Faigel, Eduard Sergienko, Andrey Bobkov, Nathalie Meurice, Joachim L. Petit, Alysia Polito, Thomas R. Caulfield, Erik P. Castle, John A. Copland, Debabrata Mukhopadhyay, Krishnendu Pal, Shamit K. Dutta, Huijun Luo, Thai H. Ho, Douglas F. Lake

    Quiescin sulfhydryl oxidase 1 (QSOX1) is an enzyme overexpressed by many different tumor types. QSOX1 catalyzes the formation of disulfide bonds in proteins. Because short hairpin knockdowns (KD) of QSOX1 have been shown to suppress tumor growth and invasion in vitro and in vivo , we hypothesized that chemical compounds inhibiting QSOX1 enzymatic activity would also suppress tumor growth, invasion, and metastasis. High throughput screening using a QSOX1-based enzymatic assay revealed multiple potential QSOX1 inhibitors. One of the inhibitors, known as “SBI-183,” suppresses tumor cell growth in a Matrigel-based spheroid assay and inhibits invasion in a modified Boyden chamber, but does not affect viability of nonmalignant cells. Oral administration of SBI-183 inhibits tumor growth in 2 independent human xenograft mouse models of renal cell carcinoma. We conclude that SBI-183 warrants further exploration as a useful tool for understanding QSOX1 biology and as a potential novel anticancer agent in tumors that overexpress QSOX1.

    更新日期:2020-01-04
  • Synergism between ATM and PARP1 Inhibition Involves DNA Damage and Abrogating the G2 DNA Damage Checkpoint
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Joyce P.Y. Mak, Hoi Tang Ma, Randy Y.C. Poon

    PARP inhibitors have emerged as effective chemotherapeutic agents for BRCA1/BRCA2-deficient cancers. Another DNA damage response protein, ATM, is also increasingly being recognized as a target for synthetic lethality with PARP inhibitors. As ATM functions in both cell cycle arrest and DNA repair after DNA damage, how cells respond to inhibition of ATM and PARP1 is yet to be defined precisely. We found that loss of ATM function, either in an ATM-deficient background or after treatment with ATM inhibitors (KU-60019 or AZD0156), results in spontaneous DNA damage and an increase in PARylation. When PARP1 is also deleted or inhibited with inhibitors (olaparib or veliparib), the massive increase in DNA damage activates the G2 DNA damage checkpoint kinase cascade involving ATR, CHK1/2, and WEE1. Our data indicated that the role of ATM in DNA repair is critical for the synergism with PARP inhibitors. Bypass of the G2 DNA damage checkpoint in the absence of ATM functions occurs only after a delay. The relative insensitivity of PARP1-deficient cells to PARP inhibitors suggested that other PARP isoforms played a relatively minor role in comparison with PARP1 in synergism with ATMi. As deletion of PARP1 also increased sensitivity to ATM inhibitors, trapping of PARP1 on DNA may not be the only mechanism involved in the synergism between PARP1 and ATM inhibition. Collectively, these studies provide a mechanistic foundation for therapies targeting ATM and PARP1.

    更新日期:2020-01-04
  • Dipyridamole Enhances the Cytotoxicities of Trametinib against Colon Cancer Cells through Combined Targeting of HMGCS1 and MEK Pathway
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Sheng Zhou, Huanji Xu, Qiulin Tang, Hongwei Xia, Feng Bi

    Both the MAPK pathway and mevalonate (MVA) signaling pathway play an increasingly significant role in the carcinogenesis of colorectal carcinoma, whereas the cross-talk between these two pathways and its implication in targeted therapy remains unclear in colorectal carcinoma. Here, we identified that HMGCS1 (3-hydroxy-3-methylglutaryl-CoA synthase 1), the rate-limiting enzyme of the MVA pathway, is overexpressed in colon cancer tissues and positively regulates the cell proliferation, migration, and invasion of colon cancer cells. In addition, HMGCS1 could enhance the activity of pERK independent of the MVA pathway, and the suppression of HMGCS1 could completely reduce the EGF-induced proliferation of colon cancer cells. Furthermore, we found that trametinib, a MEK inhibitor, could only partially abolish the upregulation of HMGCS1 induced by EGF treatment, while combination with HMGCS1 knockdown could completely reverse the upregulation of HMGCS1 induced by EGF treatment and increase the sensitivity of colon cancer cells to trametinib. Finally, we combined trametinib and dipyridamole, a common clinically used drug that could suppress the activity of SREBF2 (sterol regulatory element-binding transcription factor 2), a transcription factor regulating HMGCS1 expression, and identified its synergistic effect in inhibiting the proliferation and survival of colon cancer cells in vitro as well as the in vivo tumorigenic potential of colon cancer cells. Together, the current data indicated that HMGCS1 may be a novel biomarker, and the combination of targeting HMGCS1 and MEK might be a promising therapeutic strategy for patients with colon cancer.

    更新日期:2020-01-04
  • Dual Inhibition of Angiopoietin-TIE2 and MET Alters the Tumor Microenvironment and Prolongs Survival in a Metastatic Model of Renal Cell Carcinoma
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    May Elbanna, Ashley R. Orillion, Nur P. Damayanti, Remi Adelaiye-Ogala, Li Shen, Kiersten Marie Miles, Sreenivasulu Chintala, Eric Ciamporcero, Swathi Ramakrishnan, Sheng-yu Ku, Karen Rex, Sean Caenepeel, Angela Coxon, Roberto Pili

    Receptor tyrosine kinase inhibitors have shown clinical benefit in clear cell renal cell carcinoma (ccRCC), but novel therapeutic strategies are needed. The angiopoietin/Tie2 and MET pathways have been implicated in tumor angiogenesis, metastases, and macrophage infiltration. In our study, we used trebananib, an angiopoietin 1/2 inhibitor, and a novel small-molecule MET kinase inhibitor in patient-derived xenograft (PDX) models of ccRCC. Our goal was to assess the ability of these compounds to alter the status of tumor-infiltrating macrophages, inhibit tumor growth and metastases, and prolong survival. Seven-week-old SCID mice were implanted subcutaneously or orthotopically with human ccRCC models. One month postimplantation, mice were treated with angiopoietin 1/2 inhibitor trebananib (AMG 386), MET kinase inhibitor, or combination. In our metastatic ccRCC PDX model, RP-R-02LM, trebananib alone, and in combination with a MET kinase inhibitor, significantly reduced lung metastases and M2 macrophage infiltration ( P = 0.0075 and P = 0.0205, respectively). Survival studies revealed that treatment of the orthotopically implanted RP-R-02LM tumors yielded a significant increase in survival in both trebananib and combination groups. In addition, resection of the subcutaneously implanted primary tumor allowed for a significant survival advantage to the combination group compared with vehicle and both single-agent groups. Our results show that the combination of trebananib with a MET kinase inhibitor significantly inhibits the spread of metastases, reduces infiltrating M2-type macrophages, and prolongs survival in our highly metastatic ccRCC PDX model, suggesting a potential use for this combination therapy in treating patients with ccRCC.

    更新日期:2020-01-04
  • Precision Chemoradiotherapy for HER2 Tumors Using Antibody Conjugates of an Auristatin Derivative with Reduced Cell Permeability
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Dina V. Hingorani, Matthew K. Doan, Maria F. Camargo, Joseph Aguilera, Seung M. Song, Donald Pizzo, Daniel J. Scanderbeg, Ezra E.W. Cohen, Andrew M. Lowy, Stephen R. Adams, Sunil J. Advani

    The most successful therapeutic strategies for locally advanced cancers continue to combine decades-old classical radiosensitizing chemotherapies with radiotherapy. Molecular targeted radiosensitizers offer the potential to improve the therapeutic ratio by increasing tumor-specific kill while minimizing drug delivery and toxicity to surrounding normal tissue. Auristatins are a potent class of anti-tubulins that sensitize cells to ionizing radiation damage and are chemically amenable to antibody conjugation. To achieve tumor-selective radiosensitization, we synthesized and tested anti-HER2 antibody–drug conjugates of two auristatin derivatives with ionizing radiation. Monomethyl auristatin E (MMAE) and monomethyl auristatin F (MMAF) were attached to the anti-HER2 antibodies trastuzumab and pertuzumab through a cleavable linker. While MMAE is cell permeable, MMAF has limited cell permeability as free drug resulting in diminished cytotoxicity and radiosensitization. However, when attached to trastuzumab or pertuzumab, MMAF was as efficacious as MMAE in blocking HER2-expressing tumor cells in G2–M. Moreover, MMAF anti-HER2 conjugates selectively killed and radiosensitized HER2-rich tumor cells. Importantly, when conjugated to targeting antibody, MMAF had the advantage of decreased bystander and off-target effects compared with MMAE. In murine xenograft models, MMAF anti-HER2 antibody conjugates had less drug accumulated in the normal tissue surrounding tumors compared with MMAE. Therapeutically, systemically injected MMAF anti-HER2 conjugates combined with focal ionizing radiation increased tumor control and improved survival of mice with HER2-rich tumor xenografts. In summary, our results demonstrate the potential of cell-impermeable radiosensitizing warheads to improve the therapeutic ratio of radiotherapy by leveraging antibody–drug conjugate technology. This article is featured in Highlights of This Issue, [p. 1][1] [1]: /lookup/volpage/19/1?iss=1

    更新日期:2020-01-04
  • Efficacy of the Antibody-Drug Conjugate W0101 in Preclinical Models of IGF-1 Receptor Overexpressing Solid Tumors
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Barbara Akla, Matthieu Broussas, Noureddine Loukili, Alain Robert, Charlotte Beau-Larvor, Martine Malissard, Nicolas Boute, Thierry Champion, Jean-Francois Haeuw, Alain Beck, Michel Perez, Cyrille Dreyfus, Mariya Pavlyuk, Eric Chetaille, Nathalie Corvaia

    The insulin-like growth factor type 1 receptor (IGF-1R) is important in tumorigenesis, and its overexpression occurs in numerous tumor tissues. To date, therapeutic approaches based on mAbs and tyrosine kinase inhibitors targeting IGF-1R have only shown clinical benefit in specific patient populations. We report a unique IGF-1R–targeted antibody–drug conjugate (ADC), W0101, designed to deliver a highly potent cytotoxic auristatin derivative selectively to IGF-1R overexpressing tumor cells. The mAb (hz208F2-4) used to prepare the ADC was selected for its specific binding properties to IGF-1R compared with the insulin receptor, and for its internalization properties. Conjugation of a novel auristatin derivative drug linker to hz208F2-4 did not alter its binding and internalization properties. W0101 induced receptor-dependent cell cytotoxicity in vitro when applied to various cell lines overexpressing IGF-1R, but it did not affect normal cells. Efficacy studies were conducted in several mouse models expressing different levels of IGF-1R to determine the sensitivity of the tumors to W0101. W0101 induced potent tumor regression in certain mouse models. Interestingly, the potency of W0101 correlated with the expression level of IGF-1R evaluated by IHC. In an MCF-7 breast cancer model with high-level IGF-1R expression, a single injection of W0101 3 mg/kg led to strong inhibition of tumor growth. W0101 provides a potential new therapeutic option for patients overexpressing IGF-1R. A first-in-human trial of W0101 is currently ongoing to address clinical safety.

    更新日期:2020-01-04
  • Minicircle DNA-Engineered CAR T Cells Suppressed Tumor Growth in Mice
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Jinsheng Han, Fei Gao, Songsong Geng, Xueshuai Ye, Tie Wang, Pingping Du, Ziqi Cai, Zexian Fu, Zhilong Zhao, Long Shi, Qingxia Li, Jianhui Cai

    Viral-based chimeric antigen receptor-engineered T (CAR T)–cell manufacturing has potential safety risks and relatively high costs. The nonviral minicircle DNA (mcDNA) is safer for patients, cheaper to produce, and may be a more suitable technique to generate CAR T cells. In this study, we produced mcDNA-based CAR T cells specifically targeting prostate stem cell antigen (PSCA; mcDNA-PSCA-CAR T cells). Our results showed that mcDNA-PSCA-CAR T cells persisted in mouse peripheral blood as long as 28 days and demonstrated more CAR T-cell infiltration, higher cytokine secretion levels, and better antitumor effects. Together, our results suggest that mcDNA-CAR can be a safe and cost-effective platform to produce CAR T cells.

    更新日期:2020-01-04
  • Role of Dimerized C16orf74 in Aggressive Pancreatic Cancer: A Novel Therapeutic Target
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Toshihiro Kushibiki, Toru Nakamura, Masumi Tsuda, Takahiro Tsuchikawa, Koji Hontani, Kazuho Inoko, Mizuna Takahashi, Toshimichi Asano, Keisuke Okamura, Soichi Murakami, Yo Kurashima, Yuma Ebihara, Takehiro Noji, Yoshitsugu Nakanishi, Kimitaka Tanaka, Nako Maishi, Katsunori Sasaki, Woong-Ryeon Park, Toshiaki Shichinohe, Kyoko Hida, Shinya Tanaka, Satoshi Hirano

    Over the past 30 years, the therapeutic outcome for pancreatic ductal adenocarcinoma (PDAC) has remained stagnant due to the lack of effective treatments. We performed a genome-wide analysis to identify novel therapeutic targets for PDAC. Our analysis showed that Homo sapiens chromosome 16 open reading frame 74 (C16orf74) was upregulated in most patients with PDAC and associated with poor prognosis. Previously, we demonstrated that C16orf74 interacts with the catalytic subunit alpha of protein phosphatase 3 and plays an important role in PDAC invasion. However, the pathophysiologic function of C16orf74 is still unclear. In this study, through the analysis of C16orf74 interaction, we demonstrate a new strategy to inhibit the growth and invasion of PDAC. C16orf74 exists in the homodimer form under the cell membrane and binds integrin αVβ3 and is also involved in invasion by activating Rho family (Rac1) and MMP2. Considering that this dimeric form was found to be involved in the function of C16orf74, we designed an 11R-DB (dimer block) cell-permeable dominant-negative peptide that inhibits the dimer form of C16orf74. 11R-DB suppressed invasion and proliferation of PDAC cell lines by inhibiting phosphorylation of Akt and mTOR and also by inactivation of MMP2. 11R-DB also showed antitumor effects in an orthotopic xenograft model and peritoneal metastasis model. Thus, this study demonstrates that dimerized C16orf74, present in the cell membrane, is involved in pancreatic cancer invasion and proliferation. In addition, the C16orf74 dimer block cell-permeable peptide (11R-DB) has a potent therapeutic effect on PDAC in vitro and in vivo .

    更新日期:2020-01-04
  • ALDH1A1 Contributes to PARP Inhibitor Resistance via Enhancing DNA Repair in BRCA2-/- Ovarian Cancer Cells
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Lu Liu, Shurui Cai, Chunhua Han, Ananya Banerjee, Dayong Wu, Tiantian Cui, Guozhen Xie, Junran Zhang, Xiaoli Zhang, Eric McLaughlin, Ming Yin, Floor J. Backes, Arnab Chakravarti, Yanfang Zheng, Qi-En Wang

    Poly (ADP-ribose) polymerase (PARP) inhibitors (PARPi) are approved to treat recurrent ovarian cancer with BRCA1 or BRCA2 mutations, and as maintenance therapy for recurrent platinum-sensitive ovarian cancer (BRCA wild-type or mutated) after treatment with platinum. However, the acquired resistance against PARPi remains a clinical hurdle. Here, we demonstrated that PARP inhibitor (olaparib)–resistant epithelial ovarian cancer (EOC) cells exhibited an elevated aldehyde dehydrogenase (ALDH) activity, mainly contributed by increased expression of ALDH1A1 due to olaparib-induced expression of BRD4, a member of bromodomain and extraterminal (BET) family protein. We also revealed that ALDH1A1 enhanced microhomology-mediated end joining (MMEJ) activity in EOC cells with inactivated BRCA2, a key protein that promotes homologous recombination (HR) by using an intrachromosomal MMEJ reporter. Moreover, NCT-501, an ALDH1A1-selective inhibitor, can synergize with olaparib in killing EOC cells carrying BRCA2 mutation in both in vitro cell culture and the in vivo xenograft animal model. Given that MMEJ activity has been reported to be responsible for PARPi resistance in HR-deficient cells, we conclude that ALDH1A1 contributes to the resistance to PARP inhibitors via enhancing MMEJ in BRCA2−/− ovarian cancer cells. Our findings provide a novel mechanism underlying PARPi resistance in BRCA2-mutated EOC cells and suggest that inhibition of ALDH1A1 could be exploited for preventing and overcoming PARPi resistance in EOC patients carrying BRCA2 mutation.

    更新日期:2020-01-04
  • PD-1/PD-L1 Immune Checkpoint Inhibition with Radiation in Bladder Cancer: In Situ and Abscopal Effects
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Alexis Rompré-Brodeur, Surashri Shinde-Jadhav, Mina Ayoub, Ciriaco A. Piccirillo, Jan Seuntjens, Fadi Brimo, Jose Joao Mansure, Wassim Kassouf

    The combination of radiation with immune checkpoint inhibitors was reported in some cancers to have synergic effects both locally and distally. Our aim was to assess this combined therapy on both radiated and nonradiated bladder tumors and to characterize the immune landscape within the tumor microenvironment. Murine bladder cancer cells (MB49) were injected subcutaneously in both flanks of C57BL/6 mice. Mice were randomly assigned to the following treatments: placebo, anti-PD-L1 (four intraperitoneal injections over 2 weeks), radiation to right flank (10 Gy in two fractions), or radiation+anti-PD-L1. Tumor digestion, flow cytometry, and qPCR were performed. Log-rank analysis was used for statistical significance. Radiation+anti-PD-L1 group demonstrated statistically significant slower tumor growth rate both in the radiated and nonirradiated tumors ( P < 0.001). Survival curves demonstrated superior survival in the combination group compared with each treatment alone ( P = 0.02). Flow cytometry showed increased infiltration of immunosuppressive cells as well as CTL in the radiation and combination groups ( P = 0.04). Ratio of immunosuppressive cells to CTL shifted in favor of cytotoxic activity in the combination arm ( P < 0.001). The qPCR analysis revealed downregulation of immunosuppressive genes ( CCL22, IL22 , and IL13 ), as well as upregulation of markers of CTL activation ( CXCL9 , GZMA , and GZMB ) within both the radiated and distant tumors within the combination group. Combining radiation with immune checkpoint inhibitor provided better response in the radiated tumors and also the distant tumors along with a shift within the tumor microenvironment favoring cytotoxic activity. These findings demonstrate a possible abscopal effect in urothelial carcinoma with combination therapy.

    更新日期:2020-01-04
  • Blockade of Glutathione Metabolism in IDH1-Mutated Glioma
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Xiaoying Tang, Xiao Fu, Yang Liu, Di Yu, Sabrina J. Cai, Chunzhang Yang

    Mutations in genes encoding isocitrate dehydrogenases ( IDH ) 1 and 2 are common cancer-related genetic abnormalities. Malignancies with mutated IDHs exhibit similar pathogenesis, metabolic pattern, and resistance signature. However, an effective therapy against IDH1 -mutated solid tumor remains unavailable. In this study, we showed that acquisition of IDH1 mutation results in the disruption of NADP+/NADPH balance and an increased demand for glutathione (GSH) metabolism. Moreover, the nuclear factor erythroid 2–related factor 2 (Nrf2) plays a key protective role in IDH1 -mutated cells by prompting GSH synthesis and reactive oxygen species scavenging. Pharmacologic inhibition of the Nrf2/GSH pathway via brusatol administration exhibited a potent tumor suppressive effect on IDH1 -mutated cancer in vitro and in vivo . Our findings highlight a possible therapeutic strategy that could be valuable for IDH1 -mutated cancer treatment.

    更新日期:2020-01-04
  • Enzalutamide-Induced Feed-Forward Signaling Loop Promotes Therapy-Resistant Prostate Cancer Growth Providing an Exploitable Molecular Target for Jak2 Inhibitors
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Vindhya Udhane, Cristina Maranto, David T. Hoang, Lei Gu, Andrew Erickson, Savita Devi, Pooja G. Talati, Anjishnu Banerjee, Kenneth A. Iczkowski, Kenneth Jacobsohn, William A. See, Tuomas Mirtti, Deepak Kilari, Marja T. Nevalainen

    The second-generation antiandrogen, enzalutamide, is approved for castrate-resistant prostate cancer (CRPC) and targets androgen receptor (AR) activity in CRPC. Despite initial clinical activity, acquired resistance to enzalutamide arises rapidly and most patients develop terminal disease. Previous work has established Stat5 as a potent inducer of prostate cancer growth. Here, we investigated the significance of Jak2–Stat5 signaling in resistance of prostate cancer to enzalutamide. The levels of Jak2 and Stat5 mRNA, proteins and activation were evaluated in prostate cancer cells, xenograft tumors, and clinical prostate cancers before and after enzalutamide therapy. Jak2 and Stat5 were suppressed by genetic knockdown using lentiviral shRNA or pharmacologic inhibitors. Responsiveness of primary and enzalutamide-resistant prostate cancer to pharmacologic inhibitors of Jak2–Stat5 signaling was assessed in vivo in mice bearing prostate cancer xenograft tumors. Patient-derived prostate cancers were tested for responsiveness to Stat5 blockade as second-line treatment after enzalutamide ex vivo in tumor explant cultures. Enzalutamide-liganded AR induces sustained Jak2–Stat5 phosphorylation in prostate cancer leading to the formation of a positive feed-forward loop, where activated Stat5, in turn, induces Jak2 mRNA and protein levels contributing to further Jak2 activation. Mechanistically, enzalutamide-liganded AR induced Jak2 phosphorylation through a process involving Jak2-specific phosphatases. Stat5 promoted prostate cancer growth during enzalutamide treatment. Jak2–Stat5 inhibition induced death of prostate cancer cells and patient-derived prostate cancers surviving enzalutamide treatment and blocked enzalutamide-resistant tumor growth in mice. This work introduces a novel concept of a pivotal role of hyperactivated Jak2–Stat5 signaling in enzalutamide-resistant prostate cancer, which is readily targetable by Jak2 inhibitors in clinical development.

    更新日期:2020-01-04
  • Modulating TAK1 Expression Inhibits YAP and TAZ Oncogenic Functions in Pancreatic Cancer
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Raffaela Santoro, Marco Zanotto, Francesca Simionato, Camilla Zecchetto, Valeria Merz, Chiara Cavallini, Geny Piro, Fabio Sabbadini, Federico Boschi, Aldo Scarpa, Davide Melisi

    YAP and TAZ are central determinants of malignancy; however, their functions remain still undruggable. We identified TGFβ-activated kinase 1 (TAK1) as a central hub integrating the most relevant signals sustaining pancreatic cancer aggressiveness and chemoresistance. Glycogen synthase kinase (GSK)3 is known to stabilize TAK1, and its inhibition causes a reduction in TAK1 levels. Here, we hypothesized that TAK1 could sustain YAP/TAZ program, and thus, modulation of TAK1 expression through the inhibition of GSK3 could impair YAP/TAZ functions in pancreatic cancer. Differentially expressed transcripts between pancreatic cancer cells expressing scramble or TAK1 -specific shRNA were annotated for functional interrelatedness by ingenuity pathway analysis. TAK1 expression was modulated by using different GSK3 inhibitors, including LY2090314. In vivo activity of LY2090314 alone or in combination with nab-paclitaxel was evaluated in an orthotopic nude mouse model. Differential gene expression profiling revealed significant association of TAK1 expression with HIPPO and ubiquitination pathways. We measured a significant downregulation of YAP/TAZ and their regulated genes in shTAK1 cells. TAK1 prevented YAP/TAZ proteasomal degradation in a kinase independent manner, through a complex with TRAF6, thereby fostering their K63-ubiquitination versus K48-ubiquitination. Pharmacologic modulation of TAK1 by using GSK3 inhibitors significantly decreased YAP/TAZ levels and suppressed their target genes and oncogenic functions. In vivo , LY2090314 plus nab-paclitaxel significantly prolonged mice survival duration. Our study demonstrates a unique role for TAK1 in controlling YAP/TAZ in pancreatic cancer. LY2090314 is a novel agent that warrants further clinical development in combination with nab-paclitaxel for the treatment of pancreatic cancer.

    更新日期:2020-01-04
  • Targeting Histone Chaperone FACT Complex Overcomes 5-Fluorouracil Resistance in Colon Cancer
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Heyu Song, Jiping Zeng, Shrabasti Roychoudhury, Pranjal Biswas, Bhopal Mohapatra, Sutapa Ray, Kayvon Dowlatshahi, Jing Wang, Vimla Band, Geoffrey Talmon, Kishor K. Bhakat

    Fluorouracil (5-FU) remains a first-line chemotherapeutic agent for colorectal cancer. However, a subset of colorectal cancer patients who have defective mismatch-repair (dMMR) pathway show resistance to 5-FU. Here, we demonstrate that the efficacy of 5-FU in dMMR colorectal cancer cells is largely dependent on the DNA base excision repair (BER) pathway. Downregulation of APE1, a key enzyme in the BER pathway, decreases IC50 of 5-FU in dMMR colorectal cancer cells by 10-fold. Furthermore, we discover that the facilitates chromatin transcription (FACT) complex facilitates 5-FU repair in DNA via promoting the recruitment and acetylation of APE1 (AcAPE1) to damage sites in chromatin. Downregulation of FACT affects 5-FU damage repair in DNA and sensitizes dMMR colorectal cancer cells to 5-FU. Targeting the FACT complex with curaxins, a class of small molecules, significantly improves the 5-FU efficacy in dMMR colorectal cancer in vitro (∼50-fold decrease in IC50) and in vivo xenograft models. We show that primary tumor tissues of colorectal cancer patients have higher FACT and AcAPE1 levels compared with adjacent nontumor tissues. Additionally, there is a strong clinical correlation of FACT and AcAPE1 levels with colorectal cancer patients' response to chemotherapy. Together, our study demonstrates that targeting FACT with curaxins is a promising strategy to overcome 5-FU resistance in dMMR colorectal cancer patients.

    更新日期:2020-01-04
  • Synergy between EphA2-ILs-DTXp, a Novel EphA2-Targeted Nanoliposomal Taxane, and PD-1 Inhibitors in Preclinical Tumor Models
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    Walid S. Kamoun, Anne-Sophie Dugast, James J. Suchy, Stephanie Grabow, Ross B. Fulton, James F. Sampson, Lia Luus, Michael Santiago, Alexander Koshkaryev, Gang Sun, Vasileios Askoxylakis, Eric Tam, Zhaohua Richard Huang, Daryl C. Drummond, Andrew J. Sawyer

    Combinations of chemotherapy with immunotherapy have seen recent clinical success, including two approvals of anti–PD-1/L1 agents in combination with taxane-based chemotherapy in non–small cell lung cancer and triple-negative breast cancer. Here, we present a study on the combination activity and mechanistic rationale of a novel EphA2-targeted liposomal taxane (EphA2-ILs-DTXp) and anti–PD-1. This combination was highly active in mouse syngeneic tumor models, with complete responses observed in 3 of 5 models. In the EMT-6 tumor model, combination of EphA2-ILs-DTXp with anti–PD-1 resulted in a 60% complete response rate, with durable responses that were resistant to rechallenge. These responses were not observed in the absence of CD8+ T cells. Characterization of the immune infiltrates in EMT-6 tumors reveals increased CD8+ T cells, increased CD8+ IFNγ+ CTLs, and an increased CD8/regulatory T-cell (Treg) ratio. These immunomodulatory effects were not observed in mice treated with a combination of docetaxel and anti–PD-1. Pharmacokinetic analysis revealed that the AUC of docetaxel was increased 15 times, from 52.1 to 785 ng/mL/hour, when delivered by EphA2-ILs-DTXp. A dose reduction study of EphA2-ILs-DTXp showed a dose–response relationship for both tumor growth inhibition and the CD8/Treg ratio. Our data indicate that synergism between docetaxel and anti–PD-1 is achievable with nanoliposomal delivery.

    更新日期:2020-01-04
  • Acknowledgment to Reviewers
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2020-01-01
    American Association for Cancer Research

    The Molecular Cancer Therapeutics editors wish to acknowledge with sincere appreciation the assistance of the following reviewers who have generously contributed their time and effort during the past year[1][1] in the appraisal of manuscripts. These reviewers have been enormously helpful in

    更新日期:2020-01-04
  • Predictive and Pharmacodynamic Biomarkers of Response to the Phosphatidylinositol 3-Kinase Inhibitor Taselisib in Breast Cancer Preclinical Models
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-18
    Heather M. Moore, Heidi M. Savage, Carol O’Brien, Wei Zhou, Ethan S. Sokol, Michael E. Goldberg, Ciara Metcalfe, Lori S. Friedman, Mark R. Lackner, Timothy R. Wilson

    The PI3K signaling pathway serves as a central node in regulating cell survival, proliferation, and metabolism. PIK3CA , the gene encoding the PI3K catalytic subunit p110-alpha, is commonly altered in breast cancer resulting in the constitutive activation of the PI3K pathway. Using an unbiased cell line screening approach, we tested the sensitivity of breast cancer cell lines to taselisib, a potent PI3K inhibitor, and correlated sensitivity with key biomarkers ( PIK3CA , HER2, PTEN, and ESR1 ). We further assessed how taselisib modulates downstream signaling in the different genomic backgrounds that occur within breast cancer. We found that sensitivity to taselisib correlated with the presence of PIK3CA mutations, but was independent of HER2 status. We further showed that HER2-amplified/ PIK3CA wild-type cell lines are not as sensitive to taselisib when compared with HER2-amplified/ PIK3CA -mutant cell lines. In a PIK3CA- mutant/PTEN null background, PI3K downstream signaling rebounded in the presence of taselisib correlating with decreased sensitivity at later time points. Finally, we observed that PIK3CA mutations cooccurred with mutations in the estrogen receptor (ER; ESR1 ) in metastatic tumors from patients with ER+ breast cancer. However, the cooccurrence of an ESR1 mutation with a PIK3CA mutation did not affect response to taselisib in a single agent setting or in combination with fulvestrant. In summary, these data suggest that development of taselisib in breast cancer should occur in a PIK3CA -mutant setting with cotreatments determined by the specific subtypes under investigation.

    更新日期:2020-01-02
  • Pralatrexate in Combination with Oxaliplatin in Advanced Esophagogastric Cancer: A Phase II Trial with Predictive Molecular Correlates
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-12
    Usha Malhotra, Sarbajit Mukherjee, Christos Fountzilas, Patrick Boland, Austin Miller, Santosh Patnaik, Kristopher Attwood, Sai Yendamuri, Araba Adjei, Eric Kannisto, Mateusz Opyrchal, Peter Bushunow, Peter Loud, Renuka Iyer, Nikhil Khushalani

    The purpose of our study was to evaluate the efficacy of a combination of pralatrexate plus oxaliplatin in advanced esophagogastric cancer (EGC), analyze the impact of polymorphisms in folate metabolism pathway genes on toxicity and efficacy of pralatrexate, and to evaluate microRNA profile of tumor epithelium as a predictive biomarker. This was a two-stage trial with a safety lead in cohort and a primary endpoint of overall response rate (ORR). Patients received biweekly intravenous oxaliplatin (85 mg/m2) and pralatrexate (Dose level 1 [D1], 120 mg/m2; dose level-1 [D-1] 100 mg/m2). Single-nucleotide polymorphisms (SNP) in genes encoding proteins involved in pralatrexate metabolism were evaluated in germline DNA. microRNA profiling of the tumor epithelium was performed. ORR was 26%. Dose-limiting toxicities were observed in 2 of 4 patients at D1 and none at D-1. The T>C polymorphism in DHFR rs11951910 was significantly associated with lower progression-free survival (PFS; P ≤ 0.01), whereas the presence of the SLC19A1 rs2838957 G>A polymorphism was associated with improved PFS ( P = 0.02). Presence of the GGH rs3780130 A>T and SLC19A1 rs1051266 G>A polymorphisms were significantly associated with better overall survival (OS; P = 0.01), whereas GGH rs7010484 T>C polymorphism was associated significantly with reduced OS ( P = 0.04). There was no correlation between epithelial microRNA expression profile with disease progression or response. We conclude that the combination of oxaliplatin and pralatrexate is safe, is well tolerated, and has modest efficacy in advanced EGC. Pharmacogenomic analysis may be relevant to the use of pralatrexate in combination with platinum agents.

    更新日期:2020-01-02
  • Development of AO-176, a Next Generation Humanized Anti-CD47 Antibody With Novel Anti-Cancer Properties and Negligible Red Blood Cell Binding
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-26
    Robyn J Puro, Myriam N Bouchlaka, Ronald R Hiebsch, Benjamin J Capoccia, Michael J Donio, Pamela T Manning, William A Frazier, Robert W Karr, Daniel S Pereira

    Inhibitors of adaptive immune checkpoints have shown promise as cancer treatments. CD47 is an innate immune checkpoint receptor broadly expressed on normal tissues and overexpressed on many tumors. Binding of tumor CD47 to signal regulatory protein alpha (SIRPα) on macrophages and dendritic cells triggers a "don't eat me" signal that inhibits phagocytosis enabling escape of innate immune surveillance. Blocking CD47/SIRPα interaction promotes phagocytosis reducing tumor burden in numerous xenograft and syngeneic animal models. We have developed a next generation humanized anti-CD47 antibody, AO-176, that not only blocks the CD47/SIRPα interaction to induce tumor cell phagocytosis, but also induces tumor cytotoxicity in hematologic and solid human tumor cell lines, but not normal non-cancerous cells, by a cell autonomous mechanism (not ADCC). AO-176 also binds preferentially to tumor versus many normal cell types. In particular, AO-176 binds negligibly to RBCs in contrast to tumor cells, even at high concentrations up to 200 μg/ml and does not agglutinate RBCs up to 1 mg/ml in vitro. These properties are expected not only to decrease the antigen sink, but also to minimize on-target clinical adverse effects observed following treatment with other reported RBC-binding anti-CD47 antibodies. When tested in cynomolgus monkeys, AO-176 was well tolerated with no adverse effects. Lastly, we show that AO-176 demonstrates dose-dependent anti-tumor activity in tumor xenograft models. Taken together, the unique properties and anti-tumor activity of our next generation anti-CD47 antibody, AO-176, distinguishes it from other CD47/SIRPα axis targeting agents in clinical development.

    更新日期:2019-12-27
  • Genomic and expression analyses define MUC17 and PCNX1 as predictors of chemotherapy response in breast cancer
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-26
    Waleed S Al Amri, Lisa M Allinson, Diana E Baxter, Sandra M Bell, Andrew M. Hanby, Stacey J Jones, Abeer M Shaaban, Lucy F Stead, Eldo T Verghese, Thomas A Hughes

    Poor prognosis breast cancers are treated with cytotoxic chemotherapy, but often without any guidance from therapy predictive markers since universally-accepted markers are not currently available. Treatment failure, in the form of recurrences, is relatively common. We aimed to identify chemotherapy predictive markers and resistance pathways in breast cancer. Our hypothesis was that tumour cells remaining after neoadjuvant chemotherapy (NAC) contain somatic variants causing therapy resistance, while variants present pre-NAC but lost post-NAC cause sensitivity. Whole exome sequencing was performed on matched pre- and post-NAC cancer cells, which were isolated by laser microdissection, from 6 cancer cases, and somatic variants selected for or against by NAC were identified. Somatic variant diversity was significantly reduced after therapy (p<0.05). MUC17 variants were identified in 3 tumours and were selected against by NAC in each case, while PCNX1 variants were identified in 2 tumours and were selected for in both cases, implicating the function of these genes in defining chemoresponse. In vitro knock-down of MUC17 or PCNX1 was associated with significantly increased or decreased chemotherapy sensitivity respectively (p<0.05), further supporting their roles in chemotherapy response. Expression was tested for predictive value in two independent cohorts of chemotherapy-treated breast cancers (n=53, n=303). Kaplan-Meier analyses revealed that low MUC17 expression was significantly associated with longer survival after chemotherapy, while low PCNX1 was significantly associated with reduced survival. We concluded that therapy-driven selection of somatic variants allows identification of chemotherapy response genes. With respect to MUC17 and PCNX1, therapy-driven selection acting on somatic variants, in vitro knock-down data concerning drug sensitivity, and survival analysis of expression levels in patient cohorts all define the genes as mediators of and predictive markers for chemotherapy response in breast cancer.

    更新日期:2019-12-27
  • Targeting phosphorylation of Y-box binding protein YBX1 by TAS0612 and everolimus in overcoming antiestrogen resistance
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-26
    Tomohiro Shibata, Kosuke Watari, Akihiko Kawahara, Tomoya Sudo, Satoshi Hattori, Yuichi Murakami, Hiroto Izumi, Junji Itou, Masakazu Toi, Jun Akiba, Yoshito Akagi, Maki Tanaka, Michihiko Kuwano , Mayumi Ono

    Nuclear expression of Y-box binding protein (YBX1) is closely correlated with clinical poor outcomes and drug resistance in breast cancer. Nuclear translocation of YBX1 is facilitated by YBX1 phosphorylation at serine 102 by AKT, p70S6K and p90RSK, and the phosphorylated YBX1 (pYBX1) promotes expression of genes related to drug resistance and cell growth. A forthcoming problem to be addressed is whether targeting the phosphorylation of YBX1 overcomes antiestrogen resistance by progressive breast cancer. Here we found that increased expression of pYBX1 was accompanied by acquired resistance to antiestrogens, fulvestrant and tamoxifen. Forced expression of YBX1/S102E, a constitutive phosphorylated form, resulted in acquired resistance to fulvestrant. Inversely, YBX1 silencing specifically overcame antiestrogen resistance. Further, treatment with everolimus, an mTORC1 inhibitor, or TAS0612, a novel multikinase inhibitor of AKT, p70S6K and p90RSK, suppressed YBX1 phosphorylation and overcame antiestrogen resistance in vitro and in vivo. IHC analysis revealed that expression of pYBX1 and YBX1 was augmented in patients who experienced recurrence during treatment with adjuvant endocrine therapies. Furthermore, pYBX1 was highly expressed in patients with triple-negative breast cancer as compared with other subtypes. TAS0612 also demonstrated antitumor effect against triple-negative breast cancer in vivo. Taken together our findings suggest that pYBX1 represents a potential therapeutic target for treatment of antiestrogen resistant and progressive breast cancer.

    更新日期:2019-12-27
  • Pin1 inhibition sensitizes chemotherapy in gastric cancer cells by targeting stem-cell like traits and multiple biomarkers
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-26
    Zhen-Zhen Zhang, Wei-Xing Yu, Min Zheng, Xin-Hua Liao, Ji-Chuang Wang, Da-Yun Yang, Wen-Xian Lu, Long Wang, Sheng Zhang, He-Kun Liu, Xiao Zhen Zhou, Kun Ping Lu

    Gastric cancer is the third leading cause of cancer-related death worldwide. Diffuse type gastric cancer has the worst prognosis due to notorious resistance to chemotherapy and enrichment of cancer stem-like cells (CSCs) associated with the epithelial-mesenchymal transition (EMT). The unique proline isomerase Pin1 is a common regulator of oncogenic signaling networks and is important for gastric cancer development. However, little is known about its roles in CSCs and drug resistance in gastric cancer. In this manuscript, we demonstrate that Pin1 overexpression is closely correlated with advanced tumor stages, poor chemo-response and shorter recurrence-free survival in diffuse type gastric cancer in human patients. Furthermore, shRNA-mediated genetic or all-trans retinoic acid-mediated pharmaceutical inhibition of Pin1 in multiple human gastric cancer cells potently suppresses the EMT, cell migration and invasion, and lung metastasis. Moreover, Pin1 genetic or pharmaceutical inhibition potently eliminates gastric CSCs and suppresses their self-renewal and tumorigenicity in vitro and in vivo. Consistent with these phenotypes are that Pin1 biochemically targets multiple signaling molecules and biomarkers in EMT and CSCs and that genetic and pharmaceutical Pin1 inhibition functionally and drastically enhances the sensitivity of gastric cancer to multiple chemotherapy drugs in vitro and in vivo. These results demonstrate that Pin1 inhibition sensitizes chemotherapy in gastric cancer cells by targeting CSCs, and suggest that Pin1 inhibitors may be used to overcome drug resistance in gastric cancer.

    更新日期:2019-12-27
  • Site-specific PEGylation of anti-mesothelin recombinant immunotoxins increases half-life and anti-tumor activity
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-23
    Zeliang Zheng, Ryuhei Okada, Hisataka Kobayashi, Tadanobu Nagaya, Junxia Wei, Qi Zhou, Fred Lee, Tapan K. Bera, Yun Gao, William Kuhlman, Chin-Hsien Tai, Ira Pastan

    Recombinant Immunotoxins (RITs) are chimeric proteins containing an Fv that binds to tumor cells, fused to a fragment of Pseudomonas exotoxin (PE) that kills the cell. Their efficacy is limited by their short half-life in the circulation. Chemical modification with polyethylene glycol (PEG) is a well-established method to extend the half-lives of biologics. Our goal was to engineer RITs with an increase in half-life and high cytotoxic activity. We introduced single cysteines at different locations in five anti-mesothelin RITs and employed site-specific PEGylation to conjugate them to 20kD PEG. Because our previous PEGylation method using β-mercapto-ethanol reduction, gave poor yields of PEG-modified protein, we employed a new method using TCEP to reduce the protein, and could PEGylate RITs at ~90% efficiency. The new proteins retained 19-65% of cytotoxic activity. Although all proteins are modified with the same PEG, the radius of hydration varies from 5.2 to 7.1 showing PEG location has a large effect on protein shape. The RIT with the smallest radius of hydration has the highest cytotoxic activity. The PEGylated RITs have a 10-30-fold increase in half-life which is related to the increase in hydrodynamic size. Biodistribution experiments indicate that the long half-life is due to delayed uptake by the kidney. Anti-tumor experiments show that several PEG-RITs are much more active than unmodified RIT and the PEG location greatly affects anti-tumor activity. We conclude that PEGylation is a useful approach to improve the half-life and anti-tumor activity of RITs.

    更新日期:2019-12-25
  • High-Throughput Generation of Bispecific Binding Proteins by Sortase A-Mediated Coupling for Direct Functional Screening in Cell Culture
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-23
    Fabio Andres, Martin Schwill, Ykelien L. Boersma, Andreas Plückthun

    High-throughput construction of multivalent binders and subsequent screening for biological activity represents a fundamental challenge: a linear increase of monovalent components translates to the square of possible bivalent combinations. Even high-efficiency cloning and expression methods become limiting when thousands of bispecific binders need to be screened for activity. In this study, we present an in vitro method for the efficient production of flexibly linked bispecific binding agents from individually expressed and purified monovalent binders. We established a sortase A-mediated coupling reaction to generate bispecific Designed Ankyrin Repeat Proteins (DARPins), with an optimized reaction maximizing the bivalent coupling product with low levels of monovalent side-products. These one-pot reaction mixtures could be used directly, without further purification, in cell-based assays. We generated a matrix of 441 different bispecific DARPins against the extracellular domains of the cancer-associated receptors EGFR, ErbB2, ErbB3, ErbB4, EpCAM and c-MET and screened on two different ErbB2-positive cancer cells lines for growth-inhibitory effects. We identified not only known but also novel biologically active biparatopic DARPins. Furthermore, we found that the cancer cell lines respond in a highly reproducible and defined manner to the treatment with the 441 different bivalent binding agents. The generated response profiles can thus be used for functional characterization of cell lines since they are strongly related to the cell line-specific surface receptor landscape. Thus, our method not only represents a robust tool for screening and lead identification of bispecific binding agents, but additionally offers an orthogonal approach for the functional characterization of cancer cell lines.

    更新日期:2019-12-25
  • Toxicology and Pharmacokinetic Studies in Mice and Non-Human Primates of the Non-Toxic, Efficient, Targeted Hexameric FasL: CTLA4-FasL
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-23
    Efi Makdasi, Shira Amsili, Alexandra Aronin, Tatyana B. Prigozhina, Kobi Tzdaka, Yosi Meir Gozlan, Liat Ben Gigi -Tamir, Jitka Yehudith Sagiv, Fanny Shkedy, Noam Shani, Mark L. Tykocinski, Michal Dranitzki Elhalel

    CTLA4-FasL, a homo-hexameric signal converter protein, is capable of inducing ro-bust apoptosis in malignant cells of the B cell lineage expressing its cognate B7 and Fas targets, while sparing non-malignant ones. This fusion protein's striking pro-apoptotic efficacy stems from its complementary abilities to coordinately activate apoptotic signals and abrogate anti-apoptotic ones. A limiting factor in translating FasL or Fas receptor agonists into the clinic has been lethal hepatotoxicity. Here we establish CTLA4-FasL's in vivo efficacy in multiple murine and xenograft models, for both systemic and subcutaneous tumors. Significantly, GLP toxicology studies in mice indicate that CTLA4-FasL given repeatedly at doses up to five times the effective dose were well-tolerated and resulted in no significant adverse events. An equivalent single dose of CTLA4-FasL administered to non-human primates was also well-tolerated, albeit with a moderate dose-dependent leukopenia that was completely re-versible. Interestingly, monkey PBMC's were more sensitive to CTLA4-FasL-induced apoptosis when tested in vitro. In both species, there was short-term elevation in serum levels of IL6, IL2 and IFNγ, although this was not associated with clinical signs of pro-inflammatory cytokine release, and further, this cytokine elevation could be com-pletely prevented by dexamethasone pre-medication. Liver toxicity was not observed in either species, as confirmed by serum liver enzyme levels and histopathologic as-sessment. In conclusion, CTLA4-FasL emerges from animal model studies as an effec-tive and safe agent for targeted FasL-mediated treatment of B7-expressing aggressive B cell lymphomas.

    更新日期:2019-12-25
  • Targeting AXL and the DNA damage response pathway as a novel therapeutic strategy in melanoma
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-23
    Karine Flem-Karlsen, Erin McFadden, Nasrin Omar, Mads H Haugen, Geir Frode Øy, Truls Ryder, Hans Petter Gullestad, Robert Hermann, Gunhild Mari Mælandsmo, Vivi Ann Flørenes

    Receptor tyrosine kinase AXL is found upregulated in various types of cancer, including melanoma, and correlates with an aggressive cancer phenotype, inducing cell proliferation and epithelial-to-mesenchymal transition. Additionally, AXL has recently been linked to chemotherapy resistance and inhibition of AXL is found to increase DNA damage and reduce expression of DNA repair proteins. In light of this, we aimed to investigate if targeting AXL together with DNA damage response proteins would be therapeutically beneficial. Using melanoma cell lines, we observed that combined reduction of AXL and CHK1/CHK2 signaling decreased proliferation, deregulated cell cycle progression, increased apoptosis and reduced expression of DNA damage response proteins. Enhanced therapeutic effect of combined- as compared to mono-treatments was further observed in a patient-derived xenograft model and, of particular interest, when applying a three-dimensional ex vivo spheroid drug-sensitivity assay on tumor cells harvested directly from 27 patients with melanoma lymph node metastases. Together, these results indicate that targeting AXL together with the DNA damage response pathway could be a promising treatment strategy in melanoma and that further investigations in patient groups lacking treatment alternatives should be pursued.

    更新日期:2019-12-25
  • Monoclonal Antibody Targeting Sialyl-di-Lewisa - Containing Internalizing and non-Internalizing Glycoproteins with Cancer Immunotherapy Development Potential
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-23
    Silvana T Tivadar, Richard S McIntosh, Jia Xin Chua, Robert Moss, Tina Parsons, Abed M. Zaitoun, Srinivasan Madhusudan, Lindy G Durrant, Mireille Vankemmelbeke

    Tumor glycans constitute attractive targets for therapeutic antibodies. The sialylated glycocalyx plays a prominent role in cancer progression and immune evasion. Here we describe the characterization of the monoclonal antibody, FG129, which targets tumor-associated sialylated glycan and demonstrate its potential for multimodal cancer therapy. FG129, obtained through BALB/c mouse immunizations with liposomes containing membrane glycan extracts from the colorectal cancer cell line LS180, is a mIgG1ƙ, that targets sialyl-di-Lewisa-containing glycoproteins. FG129, as well as its chimeric human IgG1 variant, CH129, bind with nanomolar functional affinity to a range of colorectal, pancreatic and gastric cancer cell lines. FG129 targets 74% (135/182) of pancreatic, 50% (46/92) of gastric, 36% (100/281) of colorectal, 27% (89/327) of ovarian and 21% (42/201) of non-small cell lung cancers, by immunohistochemistry. In our pancreatic cancer cohort, high FG129 glyco-epitope expression was significantly associated with poor prognosis (p=0.004). Crucially, the glyco-epitope displays limited normal tissue distribution, with FG129 binding weakly to a small percentage of cells within gallbladder, ileum, liver, oesophagus, pancreas, and thyroid tissues. Owing to glyco-epitope internalization, we validated payload delivery by CH129 through monomethyl auristatin E (MMAE) or maytansinoid (DM1 and DM4) conjugation. All three CH129 drug conjugates killed high-binding colorectal and pancreatic cancer cell lines with (sub)nanomolar potency, coinciding with significant in vivo xenograft tumor control by CH129-vcMMAE. CH129, with its restricted normal tissue distribution, avid tumor binding and efficient payload delivery, is a promising candidate for the treatment of sialyl-di-Lewisa expressing solid tumors, as an ADC or as an alternative cancer immunotherapy modality.

    更新日期:2019-12-25
  • Phosphodiesterase 3A represents a therapeutic target that drives stem cell-like property and metastasis in breast cancer
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-23
    Na Hao, Wenzhi Shen, Renle Du, Shan Jiang, Junyong Zhu, Yanan Chen, Chongbiao Huang, Yi Shi, Rong Xiang, Yunping Luo

    Considerable evidence suggests that as breast cancer progresses, genetic and epigenetic mechanisms contribute to the emergence of self-renewing cells (CSCs), which may also arise as a consequence of metastasis. Although the molecular pathways that trigger stemness and metastasis are known, key molecular and mechanistic gaps in our understanding of these processes remain unclear. Here, we first screened the inflammation-associated stemness gene phosphodiesterase 3A (PDE3A) using a medium-throughput siRNA library, which was overexpressed in breast tumors and significantly correlated with clinical progression. PDE3A induced the inflammatory nuclear factor NFκB signaling pathway by suppressing cAMP/ PKA, which promotes the expression of the stem cell-marker OCT4. In addition, PDE3A also promoted the translocation of CCDC88A) from the cytoplasm to nuclei, thereby boosting the invasion-metastasis cascade in breast cancer. Most importantly, a PDE3A selective inhibitor cilostazol dramatically suppressed breast tumor growth and reduced metastasis to the lungs in xenograft breast cancer models, with minimum toxicity. Taken together, we show that PDE3A could predispose breast cancer patients to metastases by acting as a mediator of cancer stemness. PDE3A is a potential therapeutic target for advanced breast cancer.

    更新日期:2019-12-25
  • Mechanisms of entrectinib resistance in a neuroblastoma xenograft model
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-23
    Suzanne P. MacFarland, Koumudi Naraparaju, Radhika Iyer, Peng Guan, Venkatadri Kolla, Yuxuan Hu, Kai Tan, Garrett M. Brodeur

    TrkB with its ligand, brain-derived neurotrophic factor (BDNF), are overexpressed in the majority of high-risk neuroblastomas (NBs). Entrectinib is a novel pan-TRK, ALK, and ROS1 inhibitor that has shown excellent preclinical efficacy in NB xenograft models, and recently it has entered phase 1 trials in pediatric relapsed/refractory solid tumors. We examined entrectinib-resistant NB cell lines to identify mechanisms of resistance. Entrectinib-resistant cell lines were established from five NB xenografts initially sensitive to entrectinib therapy. Clonal cell lines were established in increasing concentrations of entrectinib and had >10X increase in IC50. Cell lines underwent genomic and proteomic analysis using whole-exome sequencing, RNA-Seq, and proteomic expression profiling with confirmatory RT-PCR and Western blot analysis. There was no evidence of NTRK2 (TrkB) gene mutation in any resistant cell lines. Inhibition of TrkB was maintained in all cell lines at increasing concentrations of entrectinib (target independent). PTEN pathway downregulation and ERK/MAPK pathway upregulation were demonstrated in all resistant cell lines. One of these clones also had increased IGF1R signaling, and two additional clones had increased P75 expression, which likely increased TrkB sensitivity to ligand. In conclusion, NB lines overexpressing TrkB developed resistance to entrectinib by multiple mechanisms, including activation of ERK/MAPK and downregulation of PTEN signaling. Individual cell lines also had IGF1R activation and increased P75 expression, allowing preservation of downstream TrkB signaling in the presence of entrectinib. An understanding of changes in patterns of expression can be used to inform multimodal therapy planning in using entrectinib in phase II/III trial planning.

    更新日期:2019-12-25
  • Targeting Histone Chaperone FACT Complex Overcomes 5-Fluorouracil Resistance in Colon Cancer
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-20
    Heyu Song, Jiping Zeng, Shrabasti Roychoudhury, Pranjal Biswas, Bhopal Mohapatra, Sutapa Ray, Kayvon Dowlatshahi, Jing Wang, Vimla Band, Geoffrey Talmon, Kishor K. Bhakat

    Fluorouracil (5-FU) remains a first-line chemotherapeutic agent for colorectal cancer. However, a subset of colorectal cancer patients who have defective mismatch-repair (dMMR) pathway show resistance to 5-FU. Here, we demonstrate that the efficacy of 5-FU in dMMR colorectal cancer cells is largely dependent on the DNA base excision repair (BER) pathway. Downregulation of APE1, a key enzyme in the BER pathway, decreases IC50 of 5-FU in dMMR colorectal cancer cells by 10-fold. Furthermore, we discover that the facilitates chromatin transcription (FACT) complex facilitates 5-FU repair in DNA via promoting the recruitment and acetylation of APE1 (AcAPE1) to damage sites in chromatin. Downregulation of FACT affects 5-FU damage repair in DNA and sensitizes dMMR colorectal cancer cells to 5-FU. Targeting the FACT complex with curaxins, a class of small molecules, significantly improves the 5-FU efficacy in dMMR colorectal cancer in vitro (∼50-fold decrease in IC50) and in vivo xenograft models. We show that primary tumor tissues of colorectal cancer patients have higher FACT and AcAPE1 levels compared with adjacent nontumor tissues. Additionally, there is a strong clinical correlation of FACT and AcAPE1 levels with colorectal cancer patients' response to chemotherapy. Together, our study demonstrates that targeting FACT with curaxins is a promising strategy to overcome 5-FU resistance in dMMR colorectal cancer patients.

    更新日期:2019-12-21
  • Blockade of Glutathione Metabolism in IDH1-Mutated Glioma
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-20
    Xiaoying Tang, Xiao Fu, Yang Liu, Di Yu, Sabrina J. Cai, Chunzhang Yang

    Mutations in genes encoding isocitrate dehydrogenases ( IDH ) 1 and 2 are common cancer-related genetic abnormalities. Malignancies with mutated IDHs exhibit similar pathogenesis, metabolic pattern, and resistance signature. However, an effective therapy against IDH1 -mutated solid tumor remains unavailable. In this study, we showed that acquisition of IDH1 mutation results in the disruption of NADP+/NADPH balance and an increased demand for glutathione (GSH) metabolism. Moreover, the nuclear factor erythroid 2–related factor 2 (Nrf2) plays a key protective role in IDH1 -mutated cells by prompting GSH synthesis and reactive oxygen species scavenging. Pharmacologic inhibition of the Nrf2/GSH pathway via brusatol administration exhibited a potent tumor suppressive effect on IDH1 -mutated cancer in vitro and in vivo . Our findings highlight a possible therapeutic strategy that could be valuable for IDH1 -mutated cancer treatment.

    更新日期:2019-12-21
  • Synergism between ATM and PARP1 Inhibition Involves DNA Damage and Abrogating the G2 DNA Damage Checkpoint
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-18
    Joyce P.Y. Mak, Hoi Tang Ma, Randy Y.C. Poon

    PARP inhibitors have emerged as effective chemotherapeutic agents for BRCA1/BRCA2-deficient cancers. Another DNA damage response protein, ATM, is also increasingly being recognized as a target for synthetic lethality with PARP inhibitors. As ATM functions in both cell cycle arrest and DNA repair after DNA damage, how cells respond to inhibition of ATM and PARP1 is yet to be defined precisely. We found that loss of ATM function, either in an ATM-deficient background or after treatment with ATM inhibitors (KU-60019 or AZD0156), results in spontaneous DNA damage and an increase in PARylation. When PARP1 is also deleted or inhibited with inhibitors (olaparib or veliparib), the massive increase in DNA damage activates the G2 DNA damage checkpoint kinase cascade involving ATR, CHK1/2, and WEE1. Our data indicated that the role of ATM in DNA repair is critical for the synergism with PARP inhibitors. Bypass of the G2 DNA damage checkpoint in the absence of ATM functions occurs only after a delay. The relative insensitivity of PARP1-deficient cells to PARP inhibitors suggested that other PARP isoforms played a relatively minor role in comparison with PARP1 in synergism with ATMi. As deletion of PARP1 also increased sensitivity to ATM inhibitors, trapping of PARP1 on DNA may not be the only mechanism involved in the synergism between PARP1 and ATM inhibition. Collectively, these studies provide a mechanistic foundation for therapies targeting ATM and PARP1.

    更新日期:2019-12-19
  • Inhibition of Autotaxin with GLPG1690 Increases the Efficacy of Radiotherapy and Chemotherapy in a Mouse Model of Breast Cancer
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-18
    Xiaoyun Tang, Melinda Wuest, Matthew G.K. Benesch, Jennifer Dufour, YuanYuan Zhao, Jonathan M. Curtis, Alain Monjardet, Bertrand Heckmann, David Murray, Frank Wuest, David N. Brindley

    Autotaxin catalyzes the formation of lysophosphatidic acid, which stimulates tumor growth and metastasis and decreases the effectiveness of cancer therapies. In breast cancer, autotaxin is secreted mainly by breast adipocytes, especially when stimulated by inflammatory cytokines produced by tumors. In this work, we studied the effects of an ATX inhibitor, GLPG1690, which is in phase III clinical trials for idiopathic pulmonary fibrosis, on responses to radiotherapy and chemotherapy in a syngeneic orthotopic mouse model of breast cancer. Tumors were treated with fractionated external beam irradiation, which was optimized to decrease tumor weight by approximately 80%. Mice were also dosed twice daily with GLPG1690 or vehicle beginning at 1 day before the radiation until 4 days after radiation was completed. GLPG1690 combined with irradiation did not decrease tumor growth further compared with radiation alone. However, GLPG1690 decreased the uptake of 3′-deoxy-3′-[18F]-fluorothymidine by tumors and the percentage of Ki67-positive cells. This was also associated with increased cleaved caspase-3 and decreased Bcl-2 levels in these tumors. GLPG1690 decreased irradiation-induced C-C motif chemokine ligand-11 in tumors and levels of IL9, IL12p40, macrophage colony-stimulating factor, and IFNγ in adipose tissue adjacent to the tumor. In other experiments, mice were treated with doxorubicin every 2 days after the tumors developed. GLPG1690 acted synergistically with doxorubicin to decrease tumor growth and the percentage of Ki67-positive cells. GLPG1690 also increased 4-hydroxynonenal-protein adducts in these tumors. These results indicate that inhibiting ATX provides a promising adjuvant to improve the outcomes of radiotherapy and chemotherapy for breast cancer.

    更新日期:2019-12-19
  • A Multifunctional Therapy Approach for Cancer: Targeting Raf1- Mediated Inhibition of Cell Motility, Growth, and Interaction with the Microenvironment
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-18
    Limin Zhang, Abhinandan Pattanayak, Wenqi Li, Hyun-Kyung Ko, Graham Fowler, Ryan Gordon, Raymond Bergan

    Prostate cancer cells move from their primary site of origin, interact with a distant microenvironment, grow, and thereby cause death. It had heretofore not been possible to selectively inhibit cancer cell motility. Our group has recently shown that inhibition of intracellular activation of Raf1 with the small-molecule therapeutic KBU2046 permits, for the first time, selective inhibition of cell motility. We hypothesized that simultaneous disruption of multiple distinct functions that drive progression of prostate cancer to induce death would result in advanced disease control. Using a murine orthotopic implantation model of human prostate cancer metastasis, we demonstrate that combined treatment with KBU2046 and docetaxel retains docetaxel's antitumor action, but provides improved inhibition of metastasis, compared with monotherapy. KBU2046 does not interfere with hormone therapy, inclusive of enzalutamide-mediated inhibition of androgen receptor (AR) function and cell growth inhibition, and inclusive of the ability of castration to inhibit LNCaP-AR cell outgrowth in mice. Cell movement is necessary for osteoclast-mediated bone degradation. KBU2046 inhibits Raf1 and its downstream activation of MEK1/2 and ERK1/2 in osteoclasts, inhibiting cytoskeleton rearrangement, resorptive cavity formation, and bone destruction in vitro , with improved effects observed when the bone microenvironment is chemically modified by pretreatment with zoledronic acid. Using a murine cardiac injection model of human prostate cancer bone destruction quantified by CT, KBU2046 plus zoledronic exhibit improved inhibitory efficacy, compared with monotherapy. The combined disruption of pathways that drive cell movement, interaction with bone, and growth constitutes a multifunctional targeting strategy that provides advanced disease control.

    更新日期:2019-12-19
  • Precision Chemoradiotherapy for HER2 Tumors Using Antibody Conjugates of an Auristatin Derivative with Reduced Cell Permeability
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-18
    Dina V. Hingorani, Matthew K. Doan, Maria F. Camargo, Joseph Aguilera, Seung M. Song, Donald Pizzo, Daniel J. Scanderbeg, Ezra E.W. Cohen, Andrew M. Lowy, Stephen R. Adams, Sunil J. Advani

    The most successful therapeutic strategies for locally advanced cancers continue to combine decades-old classical radiosensitizing chemotherapies with radiotherapy. Molecular targeted radiosensitizers offer the potential to improve the therapeutic ratio by increasing tumor-specific kill while minimizing drug delivery and toxicity to surrounding normal tissue. Auristatins are a potent class of anti-tubulins that sensitize cells to ionizing radiation damage and are chemically amenable to antibody conjugation. To achieve tumor-selective radiosensitization, we synthesized and tested anti-HER2 antibody–drug conjugates of two auristatin derivatives with ionizing radiation. Monomethyl auristatin E (MMAE) and monomethyl auristatin F (MMAF) were attached to the anti-HER2 antibodies trastuzumab and pertuzumab through a cleavable linker. While MMAE is cell permeable, MMAF has limited cell permeability as free drug resulting in diminished cytotoxicity and radiosensitization. However, when attached to trastuzumab or pertuzumab, MMAF was as efficacious as MMAE in blocking HER2-expressing tumor cells in G2–M. Moreover, MMAF anti-HER2 conjugates selectively killed and radiosensitized HER2-rich tumor cells. Importantly, when conjugated to targeting antibody, MMAF had the advantage of decreased bystander and off-target effects compared with MMAE. In murine xenograft models, MMAF anti-HER2 antibody conjugates had less drug accumulated in the normal tissue surrounding tumors compared with MMAE. Therapeutically, systemically injected MMAF anti-HER2 conjugates combined with focal ionizing radiation increased tumor control and improved survival of mice with HER2-rich tumor xenografts. In summary, our results demonstrate the potential of cell-impermeable radiosensitizing warheads to improve the therapeutic ratio of radiotherapy by leveraging antibody–drug conjugate technology.

    更新日期:2019-12-19
  • Targeting protein translation by rocaglamide and didesmethylrocaglamide to treat MPNST and other sarcomas
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-17
    Long-Sheng Chang, Janet L. Oblinger, Sarah S. Burns, Jie Huang, Lawrence W. Anderson, Melinda G. Hollingshead, Rulong Shen, Li Pan, Garima Agarwal, Yulin Ren, Ryan D. Roberts, Barry R O'Keefe, A. Douglas Kinghorn, Jerry M. Collins

    Malignant peripheral nerve sheath tumors (MPNSTs) frequently overexpress eIF4F components, and the eIF4A inhibitor silvestrol potently suppresses MPNST growth. However, silvestrol has suboptimal drug-like properties, including a bulky structure, poor oral bioavailability (<2%), sensitivity to MDR1 efflux, and pulmonary toxicity in dogs. We compared 10 silvestrol-related rocaglates lacking the dioxanyl ring and found that didesmethylrocaglamide (DDR) and rocaglamide (Roc) had growth-inhibitory activity comparable to silvestrol. Structure-activity relationship analysis revealed that the dioxanyl ring present in silvestrol was dispensable for, but may enhance, cytotoxicity. Both DDR and Roc arrested MPNST cells at G2/M, increased the sub-G1 population, induced cleavage of caspases and poly(ADP-ribose) polymerase, and elevated the levels of the DNA-damage response marker γH2A.X, while decreasing the expression of AKT and ERK1/2, consistent with translation inhibition. Unlike silvestrol, DDR and Roc were not sensitive to MDR1 inhibition. Pharmacokinetic analysis confirmed that Roc had 50% oral bioavailability. Importantly, Roc, when administered intraperitoneally or orally, showed potent anti-tumor effects in an orthotopic MPNST mouse model and did not induce pulmonary toxicity in dogs as found with silvestrol. Treated tumors displayed degenerative changes and had more cleaved caspase-3-positive cells, indicative of increased apoptosis. Furthermore, Roc effectively suppressed the growth of osteosarcoma, Ewing sarcoma, and rhabdomyosarcoma cells and patient-derived xenografts. Both Roc- and DDR-treated sarcoma cells showed decreased levels of multiple oncogenic kinases, including IGF-1R. The more favorable drug-like properties of DDR and Roc and the potent anti-tumor activity of Roc suggest that these rocaglamides could become viable treatments for MPNST and other sarcomas.

    更新日期:2019-12-18
  • Aurora B kinase promotes CHIP-dependent degradation of HIF1a in prostate cancer cells
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-17
    Kuntal Biswas, Sukumar Sarkar, Neveen Said, David L. Brautigan, James M. Larner

    Hypoxia is a major factor in tumor progression and resistance to therapies, which involves elevated levels of the transcription factor HIF-1a. Here we report that prostate tumor xenografts express high levels of HIF-1a and show greatly enhanced growth in response to knockdown of the E3 ligase CHIP (C-terminus of Hsp70-interacting protein). In multiple human prostate cancer cell lines under hypoxia taxol treatment induces the degradation of HIF-1a and this response is abrogated by knockdown of CHIP, but not by E3 ligase VHL or RACK1. HIF-1a degradation is accompanied by loss-of-function, evidenced by reduced expression of HIF-1a dependent genes. CHIP-dependent HIF-1a degradation also occurs in cells arrested in mitosis by nocodazole instead of taxol. Mitotic kinase Aurora B activity is required for taxol-induced HIF-1a degradation. Purified Aurora B directly phosphorylates HIF-1a at multiple sites and these modifications enhance its polyubiquitination by CHIP in a purified reconstituted system. Our results show how activation of Aurora B promotes CHIP-dependent degradation of HIF-1a in prostate cancer cells. This new knowledge may impact the use of mitotic kinase inhibitors and open new approaches for treatment of hypoxic prostate tumors.

    更新日期:2019-12-18
  • Preclinical Evaluation of a Cabazitaxel Prodrug Using Nanoparticle Delivery for the Treatment of Taxane-Resistant Malignancies
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-17
    Binbin Xie, Jianqin wan, Xiaona chen, Weidong Han, Hangxiang Wang

    Taxane-based chemotherapeutics are clinically available as frontline treatment regimens for cervical cancer. However, drug resistance and life-threatening toxicity impair the clinical efficacy of taxanes, so more effective and less toxic therapeutic modalities are urgently needed. Cabazitaxel has attracted increasing interest due to its potential to circumvent the drug resistance by taxanes. We previously showed that tethering docosahexaenoic acid (DHA) to cabazitaxel enabled the prodrug to self-assemble into nanoparticles in water. Despite this encouraging finding, the DHA-cabazitaxel conjugate formulation requires further optimization to enhance nanoparticle retention and tumor delivery. We here integrated this conjugate into amphiphilic poly(ethylene glycol)-block-poly(D,L-lactic acid) (PEG-b-PLA) copolymers to assemble dCTX NPs. The nanoparticle abrogated P-glycoprotein-mediated resistance in cancer cells. In a docetaxel-resistant cervical tumor xenograft-bearing mouse model, the efficacy was augmented by the nanotherapy when compared with solution-based free drugs (i.e., docetaxel and cabazitaxel). Dose intensification of dCTX NPs markedly suppressed the tumor growth in this model. Detailed studies revealed that systemic toxicity was alleviated, and maximum tolerated dose of dCTX NPs was at least three times higher than that of free cabazitaxel in animals, which may enable dose increases for clinical studies. In conclusion, the new formulation addresses essential requirements in terms of the stability, safety, and translational capacity for initiating early-phase clinical trials.

    更新日期:2019-12-18
  • A Novel Salicylanilide Derivative Induces Autophagy Cell Death in Castration-Resistant Prostate Cancer via ER Stress-Activated PERK Signaling Pathway
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-12
    Chia-Ling Hsieh, Hsu-Shan Huang, Kuan-Chou Chen, Teigi Saka, Chih-Ying Chiang, Leland W.K. Chung, Shian-Ying Sung

    Metastatic castration-resistant prostate cancer (CRPC) is currently incurable. Cancer growth and progression is intimately affected by its interaction with host microenvironment. Cotargeting of the stroma and prostate cancer is therefore an emerging therapeutic strategy for metastatic CRPC. Cancer-induced osteoclastogenesis is known to contribute to CRPC bone metastasis. This study is to extend pharmacologic value of our synthesized LCC03, a derivative of 5-(2′,4′-difluorophenyl)-salicylanilide that has previously testified for its osteoclastogenesis activity, by exploring its additional cytotoxic properties and underlying mechanism in CRPC cells. LCC03 was chemically synthesized and examined for cell growth inhibition in a serial of CRPC cell lines. We demonstrated that LCC03 dose-dependently suppressed proliferation and retarded cell-cycle progression in CRPC cells. The classical autophagy features, including autophagosome formation and LC3-II conversion, were dramatically shown in LCC03-treated CRPC cells, and it was associated with the suppressed AKT/mTOR signaling pathways, a major negative regulator of autophagy. Moreover, an expanded morphology of the endoplasmic reticulum (ER), increased expression of the ER stress markers GRP78 and PERK, and eIF2α phosphorylation were observed. Blockage of autophagy and PERK pathways using small molecule inhibitors or shRNA knockdown reversed LCC03-induced autophagy and cell death, thus indicating that the PERK–eIF2α pathway contributed to the LCC03-induced autophagy. Furthermore, treatment of tumor-bearing mice with intraperitoneal administered LCC03 suppressed the growth of CRPC xenografts in mouse bone without systemic toxicity. The dual action of 5-(2′,4′-difluorophenyl)-salicylanilide on targeting both the osteoclasts and the tumor cells strongly indicates that LCC03 is a promising anticancer candidate for preventing and treating metastatic CRPC.

    更新日期:2019-12-13
  • Molecular Inhibitor of QSOX1 Suppresses Tumor Growth In Vivo
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-12
    Amber L. Fifield, Paul D. Hanavan, Douglas O. Faigel, Eduard Sergienko, Andrey Bobkov, Nathalie Meurice, Joachim L. Petit, Alysia Polito, Thomas R. Caulfield, Erik P. Castle, John A. Copland, Debabrata Mukhopadhyay, Krishnendu Pal, Shamit K. Dutta, Huijun Luo, Thai H. Ho, Douglas F. Lake

    Quiescin sulfhydryl oxidase 1 (QSOX1) is an enzyme overexpressed by many different tumor types. QSOX1 catalyzes the formation of disulfide bonds in proteins. Because short hairpin knockdowns (KD) of QSOX1 have been shown to suppress tumor growth and invasion in vitro and in vivo , we hypothesized that chemical compounds inhibiting QSOX1 enzymatic activity would also suppress tumor growth, invasion, and metastasis. High throughput screening using a QSOX1-based enzymatic assay revealed multiple potential QSOX1 inhibitors. One of the inhibitors, known as “SBI-183,” suppresses tumor cell growth in a Matrigel-based spheroid assay and inhibits invasion in a modified Boyden chamber, but does not affect viability of nonmalignant cells. Oral administration of SBI-183 inhibits tumor growth in 2 independent human xenograft mouse models of renal cell carcinoma. We conclude that SBI-183 warrants further exploration as a useful tool for understanding QSOX1 biology and as a potential novel anticancer agent in tumors that overexpress QSOX1.

    更新日期:2019-12-13
  • Clinical application of next-generation sequencing-based panel to BRAF wild-type advanced melanoma identifies key oncogenic alterations and therapeutic strategies
    Mol. Cancer Ther. (IF 4.856) Pub Date : 2019-12-11
    Changhee Park, Miso Kim, Min Jung Kim, Hyeongmin Kim, Chan-Young Ock, Bhumsuk Keam, Tae Min Kim, Dong-Wan Kim, Jong-Il Kim, Dae Seog Heo

    Molecular profiling with next-generation sequencing (NGS) has been applied in multiple solid cancers in order to discover potential therapeutic targets. Here, we describe the results of a clinical NGS panel in advanced melanoma patients. Thirty-six tumor tissues from BRAF wild-type melanoma patients at Seoul National University Hospital (SNUH) were collected and deep-sequenced using the SNUH FIRST-Cancer NGS panel to assess single nucleotide variants, small insertions/deletions, copy number variations, and structural variations to estimate tumor mutation burden (TMB). We discovered 106 oncogenic alterations and most of the patients (n = 33, 92 %) harbored at least one oncogenic alteration, including two patients who were initially diagnosed as BRAF V600E negative but were later confirmed to be positive. Altogether, 36 samples were classified into RAS/BRAF/NF1 mutant (n = 14, 39%) or triple wild-type (n = 22, 61%) melanoma subtypes. The estimated median TMB was 8.2 mutations per Mb, ranging from 0 to 146.67 mutations per Mb. Of the 36 patients, 25 (70%) had actionable alterations with currently developed drugs and 7 (19.4%) were enrolled in clinical trials with RAF inhibitor, multiple receptor tyrosine kinase inhibitor, and anti-programmed cell death-1 (PD-1) antibody. TMB tended to associate with progression-free survival (PFS) of treatment with anti-PD-1/programmed death ligand-1 antibody (hazard ratio 0.96, 95% confidence interval 0.92 - 1.00, p = 0.07). High-TMB (≥13) group was associated with longer PFS than the low-TMB group (Median 34.0 vs. 11.0 weeks, p = 0.04). Overall, the clinical use of NGS panel in advanced melanoma patients shows association with clinical outcomes and several therapeutic strategies.

    更新日期:2019-12-13
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