当前位置: X-MOL 学术 › Biol. Blood Marrow Transplant. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Transcriptome Profiling of Immune Response to Dendritic/AML Fusion Vaccine
Biology of Blood and Marrow Transplantation ( IF 5.609 ) Pub Date : 2020-01-23 , DOI: 10.1016/j.bbmt.2019.12.078
Jessica Liegel , Manoj Bhasin , German Pihan , Beena Thomas , Dina Stroopinsky , Haider Ghiassudin , Emma Logan , Adam Morin , Marzia Capelletti , Shira Orr , Cansu Tacettin , Myrna R. Nahas , Matthew Weinstock , Maryam Rahimian , Lina Bisharat , Donald Kufe , David E. Avigan , Jacalyn Rosenblatt

Introduction

In a clinical trial of patients vaccinated after chemotherapy-induced remission with patient-derived AML cells fused with autologous dendritic cells (DC/AML fusions) we demonstrated durable AML-specific immunity. 71% of patients sustained long-term remission from AML at 55-91 months (responders) while those who experienced relapse had recurrence at 2-26 months (non-responders).

Objectives

We performed genomic analysis of the marrow microenvironment to identify factors associated with durable remission after vaccination.

Methods

RNA sequencing was done using banked formalin-fixed paraffin embedded (FFPE) marrow at serial time points. Ingenuity Pathways IPA 9.0 was used to define pathways and upstream regulators. To characterize the cellular components, single cell RNA-seq was performed on fresh frozen aspirates with cell cluster annotation performed by Single Cell Wizard software and peripheral blood was used for TCR clonal diversity by NGS.

Results

Heatmaps depict significant (p-value ≤0.01 and fold change ≥2) differential gene expression both pre- and post-vaccination in responders compared to non-responders. Prior to vaccination there was inhibition of TGF-β in responders. Notable upregulated pathways in responders after vaccination (p value <0.01) were related to immune activation including NO and ROS in macrophages, IL-2, IL-15, IL-6, IL-7, IL17A, and B cell activating factor. TGF-β was also downregulated in responders post-vaccination. Key upstream transcription regulators of response pre-vaccination included inhibition of IL-4, IGF-1, VEGF, PDGFBB and TGF-β1 while post-vaccination included activation of CD3, CD28, tumor-necrosis factor and IL-1B. We also identified a highly connected cohesive network of immune system activation including these key regulators as well as GBP2, CD226 and toll-like receptor genes. In responders, increased cellular heterogeneity was present pre-vaccine, and increases in T and NK populations post-vaccine. Also, temporal changes in expression of TCR clonotypes showed a sustained increase in TCR diversity after vaccination.

Conclusion

In a cohort of AML patients treated with DC/AML fusions, we found distinct gene signatures amongst patients with long term remisison vs those with early relapse. The transcriptomes indicate a microenvironment in which TGF-β-mediated immunosuppression is inhibited after chemotherapy and in which interleukins, B cell and macrophage signaling are activated are associated with vaccine response, and have potential for combinatorial therapeutic strategies. T cell costimulation and activation are key components of response with NK cells and DC activation playing a supporting role. A durable oligoclonal T cell expansion suggests vaccine is capable of enhancing the T cell repertroie which could provide a unique opportunity to identify target antigens by TCR-epitope pairing.



中文翻译:

转录组分析树突状/ AML融合疫苗的免疫反应。

介绍

在对化疗诱导的缓解后接种自体树突状细胞(DC / AML融合)的患者源性AML细胞进行疫苗接种的患者的临床试验中,我们证明了持久的AML特异性免疫力。71%的患者在55-91个月时长期从AML缓解(应答者),而经历了复发的患者则在2-26个月复发(无应答者)。

目标

我们对骨髓微环境进行了基因组分析,以确定与疫苗接种后持久缓解相关的因素。

方法

RNA测序是在连续的时间点使用银行存的福尔马林固定石蜡包埋的(FFPE)骨髓完成的。独创性途径IPA 9.0用于定义途径和上游调节剂。为了表征细胞成分,在新鲜的冷冻吸出液上进行单细胞RNA测序,并通过Single Cell Wizard软件进行细胞簇注释,NGS将外周血用于TCR克隆多样性。

结果

热图描述了接种前和接种后与未接种者相比,显着(p值≤0.01,倍数变化≥2)差异基因表达。在接种疫苗之前,应答者中的TGF - β受到抑制。接种疫苗后应答者中显着上调的途径(p值<0.01)与免疫活化有关,包括巨噬细胞中的NO和ROS,IL-2,IL-15,IL-6,IL-7,IL17A和B细胞活化因子。接种疫苗后应答者的TGF - β也下调。响应前接种的关键上游转录调节包括IL-4,IGF-1,VEGF,PDGFBB和TGF抑制- β 1疫苗接种后包括CD3,CD28,肿瘤坏死因子和IL-1B的激活。我们还确定了高度关联的免疫系统激活的内聚网络,包括这些关键调控因子以及GBP2,CD226和toll样受体基因。在应答者中,疫苗接种前细胞异质性增加,疫苗接种后T和NK种群增加。同样,TCR克隆型表达的时间变化表明接种疫苗后TCR多样性持续增加。

结论

在接受DC / AML融合治疗的AML患者队列中,我们发现长期共存的患者与早期复发的患者具有明显的基因特征。转录组表明一种微环境,其中化学疗法后TGF - β介导的免疫抑制被抑制,并且白介素,B细胞和巨噬细胞信号传导被激活与疫苗反应相关,并且具有组合治疗策略的潜力。T细胞共刺激和激活是NK细胞反应的关键组成部分,而DC激活起辅助作用。持久的寡克隆T细胞扩增表明,疫苗能够增强T细胞全集,这可能为通过TCR表位配对鉴定靶抗原提供独特的机会。

更新日期:2020-01-23
down
wechat
bug