Heparanase 2 (Hpa2) attenuates the growth of pancreatic carcinoma
Introduction
Heparanase is an endo-beta-glucuronidase capable of cleaving heparan sulfate (HS) side chains of heparan sulfate proteoglycans (HSPG). This activity results in remodeling of the extracellular matrix (ECM) and enhances cell dissemination associated with tumor metastasis and transmigration of immune cells [1], [2], [3], [4]. Compelling evidence gathered in the last two decades tie heparanase not only with tumor metastasis but with all aspects of cancer progression namely tumor initiation, growth, metastasis, and chemoresistance [3,[5], [6], [7]]. Also, heparanase was noted to orchestrate a cross-talk between tumor cells and the tumor microenvironment [8], [9], [10], [11], [12], [13] and to support stemness [14], thus encouraging the development of heparanase inhibitors as anti-cancer drugs [3,6,7,[15], [16], [17]]. Heparanase 2 (Hpa2) is a close homolog of heparanase [18]; it lacks HS-degrading activity typical of heparanase, yet retains the high-affinity interaction with HS [19]. Unlike the intense research effort devoted to exploring the significance of heparanase in human diseases, very little attention was given to Hpa2. Notably, in head and neck cancer high levels of Hpa2 are associated with prolonged patients survival and decreased tumor cell dissemination to regional lymph nodes [19], suggesting that Hpa2 functions as a tumor suppressor. Here, we examined the role of Hpa2 in pancreatic cancer, a malignancy characterized by a dense fibrotic ECM associated with poor response to treatment and exceedingly bad prognosis. We found that patients who show high levels of Hpa2 survive longer than patients with low levels of Hpa2. Likewise, overexpression of Hpa2 in pancreatic carcinoma cells resulted in a most prominent decrease in tumor growth (orthotopic and intra-peritoneal) whereas Hpa2 silencing resulted in bigger tumors. We further found that Hpa2 elicits endoplasmic reticulum (ER) stress and increased apoptotic cell death. Importantly, ER stress induces the expression of Hpa2, thus establishing a loop that feeds itself. This loop, and the higher sensitivity of Hpa2-overexpressing cells to stress conditions, likely leads to attenuated tumor growth.
Section snippets
Hpa2 attenuates pancreatic tumor growth
To reveal the role of Hpa2 in pancreatic cancer we subjected a cohort of PDAC biopsies (Table 1) to immunostaining applying anti-Hpa2 antibody. No, or very weak staining of Hpa2 was detected in normal ducts and exocrine cells in normal pancreatic tissue adjacent to the tumor lesion (Fig. 1A, upper left panel), whereas pancreatic beta cells in Langerhans islets were stained positive. These positively-stained islets are found in the normal pancreas tissue adjacent to the tumor lesion (Fig. 1A,
Discussion
While the pro-tumorigenic properties of heparanase are well-taken [3,6,15,23], the role of Hpa2 in cancer is largely unknown. Unlike heparanase, Hpa2 staining is readily detected in the normal epithelium of the bladder, breast, gastric and ovarian tissues. Notably, Hpa2 levels are decreased substantially in the resulting carcinomas [20,[24], [25], [26]], a staining pattern typical of a tumor suppressor. Moreover, high levels of Hpa2 associate with prolonged survival of head and neck cancer
Conclusions
Our findings indicate that Hpa2 attenuates pancreatic tumor growth and highlight a feedback mechanism by which Hpa2 enhances ER stress which, in turn, induces Hpa2 expression. This results in consistent and severe ER stress, leading to increased apoptotic cell death and attenuated tumor growth. Thus, compounds designed to elicit ER stress may turn beneficial therapeutics in pancreatic cancer.
Cells, cell culture, and immunoblotting
Human Panc-01, Capan-1, and CFPAC-1 pancreatic carcinoma cells were purchased from the ATCC; mouse Panc-02 cells have been described previously [13]. Cells were grown in Dulbecco's modified Eagle's medium (Biological Industries, Beit Haemek, Israel) supplemented with 10% fetal bovine serum and antibiotics. The human cell lines were authenticated in June 2020 by the short tandem repeat (STR) profile of 15 loci plus amelogenin for sex determination (X or XY) method, according to the
Declaration of Competing Interest
The authors declare no conflict of interest.
Funding
These studies were generously supported by research grants awarded by the Israel Science Foundation (ISF grant 1021/19), and the Israel Cancer Research Fund (ICRF). I. Vlodavsky is a Research Professor of the ICRF.
Author contribution
Conception and design of the work- IV, NI,
Acquisition, analysis, or interpretation of data-YK, PS, IN, NI
Drafted the work or substantively revised it-NI, IV
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2022, Matrix BiologyCitation Excerpt :Importantly, patients that retain high levels of Hpa2 survived longer than Hpa2-low tumors [11,20,21,23,24]. Experimentally, overexpression of Hpa2 attenuated the growth of tumor xenografts, whereas Hpa2 gene silencing resulted in bigger tumors [21,23,25,26]. This is best demonstrated by gene editing of Hpa2 in pharyngeal FaDu cells applying the CRISPR technology.
Induction of heparanase 2 (Hpa2) expression by stress is mediated by ATF3
2022, Matrix BiologyCitation Excerpt :This expression pattern is characteristic of tumor suppressors. Furthermore, high levels of Hpa2 were associated with longer survival of head and neck, pancreatic and gastric cancer patients vs patients exhibiting low levels of Hpa2 [4,22,23]. Likewise, overexpression of Hpa2 in head and neck and pancreatic carcinoma cells resulted in a prominent decrease in tumor growth, mediated by lower vascular density [24], increased expression of Sox2 [25], and increased ER stress response that, once present constitutively, promote apoptotic cell death [22].
Extracellular matrix-based cancer targeting
2021, Trends in Molecular MedicineRole of heparanase 2 (Hpa2) in gastric cancer
2021, Neoplasia (United States)Citation Excerpt :Notably, metformin attenuated cell proliferation, migration, invasion, and colony formation by gastric carcinoma cells to an extent comparable with Hpa2 (Fig. 6, Suppl. Fig. 4), suggesting that these properties of Hpa2 are mediated, at least in part, by AMPK. The role of AMPK in Hpa2-mediated tumor attenuation awaits stringent confirmation in a subsequent study, employing gastric, head and neck, pancreatic [25,27,57], or other types of cancer that are affected by Hpa2 and show increased levels of AMPK phosphorylation. The mechanism by which Hpa2 promotes AMPK phosphorylation is not clear but seems to involve HS because the increased ACC phosphorylation levels in Hpa2 cells was reversed by the addition of heparin to the cell culture medium (Fig. 5C).
Heparanase 2 (Hpa2) attenuates tumor growth by inducing Sox2 expression
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Conclusions: Our findings indicate that Hpa2 attenuates pancreatic tumor growth and highlight a feedback mechanism by which Hpa2 enhances ER stress which, in turn, induces Hpa2 expression, leading to increased tumor cell death.