Periodic Reporting for period 1 - HEPINIB (Heparanase inhibition as a multifunctional targeting approach in cancer)
Periodo di rendicontazione: 2023-01-01 al 2024-12-31
Heparan sulfate (HS) is a carbohydrate of varied structure that attaches to proteins found on the cell surface and in the extracellular matrix. Emerging evidence indicates that this process is deregulated in cancer and affects cancer cell interactions with the tumour microenvironment and the immune system, promoting cancer progression and metastasis. Funded by the Marie Skłodowska-Curie Actions programme, the HEPINIB project proposes to use as a drug target the enzyme heparanase that cleaves HS and contributes to the remodelling of the extracellular matrix. The working hypothesis is that the therapeutic targeting of heparanase will work on many levels, impeding metastasis, angiogenesis and inflammation.
Cancer is a leading cause of mortality within the aging European population. Therapeutic targeting is hampered by the complexity of the disease, which includes not only molecular changes within the tumor cell itself, but also within its microenvironment. Tumor angiogenesis, tumor-stroma interactions, interactions with immune cells, with the extracellular matrix and cancer stem cell niches allow for malignant cell survival and promote metastasis, the leading cause for cancer-associated mortality. Proteins substituted with the heparin-related carbohydrate heparan sulfate (HS) are dysregulated in malignant diseases, and are known to modulate all of the aforementioned processes of tumor progression. Several functions of HS-PGs in tumor progression are modulated by the enzyme heparanase (HPSE), which is barely expressed in adults, but upregulated during tumor progression, inflammation and angiogenesis, thus constituting an excellent drug target. Indeed, HPSE, the sole HS degrading endoglycosidase, regulates multiple biological activities that enhance tumor growth, metastasis, angiogenesis and inflammation. HPSE accomplishes this by degrading HS and thereby regulating the bioavailability of heparin-binding proteins, priming the tumor microenvironment and mediating tumor-host crosstalk. We predict that therapeutic targeting of HPSE to be superior to conventional approaches, as it does not only have the potential to synchronously targeting tumor progression and metastasis at multiple levels (metastasis, angiogenesis, inflammation & immunity), but it is also be expected to have a favourable side effects profile. HEPINIB will combine leading experts of HPSE-related cancer research with non-academic partners providing a panel of innovative technologies with the aims of providing a deeper understanding of HPSE function in tumor progression, of developing novel HPSE inhibitors and delivery systems, and of trans-sectoral and interdisciplinary training of young scientists.
Cancer is a leading cause of mortality within the aging European population. Therapeutic targeting is hampered by the complexity of the disease, which includes not only molecular changes within the tumor cell itself, but also within its microenvironment. Tumor angiogenesis, tumor-stroma interactions, interactions with immune cells, with the extracellular matrix and cancer stem cell niches allow for malignant cell survival and promote metastasis, the leading cause for cancer-associated mortality. Proteins substituted with the heparin-related carbohydrate heparan sulfate (HS) are dysregulated in malignant diseases, and are known to modulate all of the aforementioned processes of tumor progression. Several functions of HS-PGs in tumor progression are modulated by the enzyme heparanase (HPSE), which is barely expressed in adults, but upregulated during tumor progression, inflammation and angiogenesis, thus constituting an excellent drug target. Indeed, HPSE, the sole HS degrading endoglycosidase, regulates multiple biological activities that enhance tumor growth, metastasis, angiogenesis and inflammation. HPSE accomplishes this by degrading HS and thereby regulating the bioavailability of heparin-binding proteins, priming the tumor microenvironment and mediating tumor-host crosstalk. We predict that therapeutic targeting of HPSE to be superior to conventional approaches, as it does not only have the potential to synchronously targeting tumor progression and metastasis at multiple levels (metastasis, angiogenesis, inflammation & immunity), but it is also be expected to have a favourable side effects profile. HEPINIB will combine leading experts of HPSE-related cancer research with non-academic partners providing a panel of innovative technologies with the aims of providing a deeper understanding of HPSE function in tumor progression, of developing novel HPSE inhibitors and delivery systems, and of trans-sectoral and interdisciplinary training of young scientists.
HEPINIB has prepared and characterized samples of heparin and low molecular weight heparins modified to eliminate the majority of the anticoagulant activity through the oxidation and reduction of the non-sulphated uronic acid residues including the glucuronic acid residue part of the binding site of heparin for antithrombin. Moreover, HEPINIB has synthesized and characterized LMW heparins specifically functionalized to be linked to structures capable of forming nanoparticles. The inhibitors have been tested in functional assays on a variety of human cancer cell lines and in vivo models, and were assayed for heparanase inhibitory activity, leading to the identification of promising lead compounds with high inhibitory activity. Of note, an impact on the cancer stem cell phenotype - related to therapeutic resistance - could be demonstrated for colon cancer, brain cancer (GMB) and in part for breast cancer in preclinical experimental models. HEPINIB has furthermore characterised heparanase expression in extracellular vesicles using advanced biophysical techniques. HEPINIB could also show that Hpa2 is expressed in the normal epithelium of the human bladder, breast, gastric and ovarian tissues and is reduced substantially in the resulting carcinomas, a staining pattern typical of a tumor suppressor. Employing gene overexpression, gene editing (CRISPR) and silencing approaches, an anti-cancerous effect of Hpa2 was demonstrated in preclinical models of head & neck, bladder, gastric, and pancreatic carcinomas, and in mouse models of human sarcomas. Hpa2 was found to regulate selected genes that promote normal differentiation, tissue homeostasis, and endoplasmic reticulum (ER) stress, resulting in antitumor, antiangiogenic and anti-inflammatory effects. Importantly, stress conditions induced the expression of Hpa2, thus establishing a feedback loop. In most cases, patients who retain high levels of Hpa2 survived longer than patients bearing Hpa2-low tumors. Our data point at translationally revevant roles for heparanase 1 and 2 in different tumor entities, which will be targeted using the consortium's inhibitors during further project implementation.
Our results indicate that the heparanase inhibitors may be effective in targeting a cancer stem cell phenotype - a property that could be exploited to overcome therapeutic resistance. Moreover, the novel role of heparanase 2 as a potential tumor suppressor in cancer is intriguing and provides another potential avenue for therapeutic targeting of cancer with heparanase-2-based drugs. HEPINIB has furthermore developed new analytical techniques (spectroscopy of GAGs, new GAG-based 3D in vitro culture models, nanoparticles for GAG delivery) with high commercialisation potential.