TGF-β inhibition combined with cytotoxic nanomedicine normalizes triple negative breast cancer microenvironment towards anti-tumor immunity

Tumor normalization strategies aim to improve tumor blood vessel functionality (i.e., perfusion) by reducing the hyper-permeability of tumor vessels or restoring compressed vessels. Despite progress in strategies to normalize the tumor microenvironment (TME), their combinatorial antitumor effects with nanomedicine and immunotherapy remain unexplored. Methods: Here, we re-purposed the TGF-β inhibitor tranilast, an approved anti-fibrotic and antihistamine drug, and combined it with Doxil nanomedicine to normalize the TME, increase perfusion and oxygenation, and enhance anti-tumor immunity. Specifically, we employed two triple-negative breast cancer (TNBC) mouse models to primarily evaluate the therapeutic and normalization effects of tranilast combined with doxorubicin and Doxil. We demonstrated the optimized normalization effects of tranilast combined with Doxil and extended our analysis to investigate the effect of TME normalization to the efficacy of immune checkpoint inhibitors. Results: Combination of tranilast with Doxil caused a pronounced reduction in extracellular matrix components and an increase in the intratumoral vessel diameter and pericyte coverage, indicators of TME normalization. These modifications resulted in a significant increase in tumor perfusion and oxygenation and enhanced treatment efficacy as indicated by the notable reduction in tumor size. Tranilast further normalized the immune TME by restoring the infiltration of T cells and increasing the fraction of T cells that migrate away from immunosuppressive cancer-associated fibroblasts. Furthermore, we found that combining tranilast with Doxil nanomedicine, significantly improved immunostimulatory M1 macrophage content in the tumorigenic tissue and improved the efficacy of the immune checkpoint blocking antibodies anti-PD-1/anti-CTLA-4. Conclusion: Combinatorial treatment of tranilast with Doxil optimizes TME normalization, improves immunostimulation and enhances the efficacy of immunotherapy.

[1]  R. Jain,et al.  Improving cancer immunotherapy using nanomedicines: progress, opportunities and challenges , 2020, Nature Reviews Clinical Oncology.

[2]  K. Toh,et al.  Dexamethasone Increases Cisplatin-Loaded Nanocarrier Delivery and Efficacy in Metastatic Breast Cancer by Normalizing the Tumor Microenvironment. , 2019, ACS nano.

[3]  R. Jain,et al.  Reprogramming the microenvironment with tumor-selective angiotensin blockers enhances cancer immunotherapy , 2019, Proceedings of the National Academy of Sciences.

[4]  R. Jain,et al.  Experimental and computational analyses reveal dynamics of tumor vessel cooption and optimal treatment strategies , 2019, Proceedings of the National Academy of Sciences.

[5]  R. Jain,et al.  Blocking CXCR4 alleviates desmoplasia, increases T-lymphocyte infiltration, and improves immunotherapy in metastatic breast cancer , 2019, Proceedings of the National Academy of Sciences.

[6]  R. Jain,et al.  Losartan treatment enhances chemotherapy efficacy and reduces ascites in ovarian cancer models by normalizing the tumor stroma , 2019, Proceedings of the National Academy of Sciences.

[7]  Federico Cappuzzo,et al.  Atezolizumab for First‐Line Treatment of Metastatic Nonsquamous NSCLC , 2018, The New England journal of medicine.

[8]  Fabian Kiessling,et al.  Tumor targeting via EPR: Strategies to enhance patient responses. , 2018, Advanced drug delivery reviews.

[9]  Triantafyllos Stylianopoulos,et al.  Reengineering the Physical Microenvironment of Tumors to Improve Drug Delivery and Efficacy: From Mathematical Modeling to Bench to Bedside. , 2018, Trends in cancer.

[10]  T. Stylianopoulos,et al.  Accumulation of mechanical forces in tumors is related to hyaluronan content and tissue stiffness , 2018, PloS one.

[11]  Inna Kuperstein,et al.  Fibroblast Heterogeneity and Immunosuppressive Environment in Human Breast Cancer. , 2018, Cancer cell.

[12]  Dai Fukumura,et al.  Enhancing cancer immunotherapy using antiangiogenics: opportunities and challenges , 2018, Nature Reviews Clinical Oncology.

[13]  Camille Stephan-Otto Attolini,et al.  TGFβ drives immune evasion in genetically reconstituted colon cancer metastasis , 2018, Nature.

[14]  R. Bourgon,et al.  TGF-β attenuates tumour response to PD-L1 blockade by contributing to exclusion of T cells , 2018, Nature.

[15]  P. Papageorgis,et al.  Sonic‐hedgehog pathway inhibition normalizes desmoplastic tumor microenvironment to improve chemo‐ and nanotherapy , 2017, Journal of controlled release : official journal of the Controlled Release Society.

[16]  T. Ishida,et al.  Metronomic chemotherapy and nanocarrier platforms. , 2017, Cancer letters.

[17]  P. Papageorgis,et al.  Tranilast-induced stress alleviation in solid tumors improves the efficacy of chemo- and nanotherapeutics in a size-independent manner , 2017, Scientific Reports.

[18]  Max Nobis,et al.  Transient tissue priming via ROCK inhibition uncouples pancreatic cancer progression, sensitivity to chemotherapy, and metastasis , 2017, Science Translational Medicine.

[19]  P. Papageorgis,et al.  Pirfenidone normalizes the tumor microenvironment to improve chemotherapy , 2017, Oncotarget.

[20]  Triantafyllos Stylianopoulos,et al.  Role of vascular normalization in benefit from metronomic chemotherapy , 2017, Proceedings of the National Academy of Sciences.

[21]  Dai Fukumura,et al.  Reengineering the Tumor Microenvironment to Alleviate Hypoxia and Overcome Cancer Heterogeneity. , 2016, Cold Spring Harbor perspectives in medicine.

[22]  V. Gkretsi,et al.  Hyaluronan-Derived Swelling of Solid Tumors, the Contribution of Collagen and Cancer Cells, and Implications for Cancer Therapy12 , 2016, Neoplasia.

[23]  T. Stylianopoulos,et al.  Biphasic modeling of brain tumor biomechanics and response to radiation treatment. , 2016, Journal of biomechanics.

[24]  R. Jain,et al.  Dual inhibition of Ang-2 and VEGF receptors normalizes tumor vasculature and prolongs survival in glioblastoma by altering macrophages , 2016, Proceedings of the National Academy of Sciences.

[25]  Jennie W. Taylor,et al.  Ang-2/VEGF bispecific antibody reprograms macrophages and resident microglia to anti-tumor phenotype and prolongs glioblastoma survival , 2016, Proceedings of the National Academy of Sciences.

[26]  R. Jain,et al.  Metformin Reduces Desmoplasia in Pancreatic Cancer by Reprogramming Stellate Cells and Tumor-Associated Macrophages , 2015, PloS one.

[27]  Triantafyllos Stylianopoulos,et al.  Design considerations for nanotherapeutics in oncology. , 2015, Nanomedicine : nanotechnology, biology, and medicine.

[28]  R. Hollingsworth,et al.  Doxil Synergizes with Cancer Immunotherapies to Enhance Antitumor Responses in Syngeneic Mouse Models , 2015, Neoplasia.

[29]  Wen Jiang,et al.  Remodeling Tumor Vasculature to Enhance Delivery of Intermediate-Sized Nanoparticles. , 2015, ACS nano.

[30]  Yi Cao,et al.  The nanomechanical signature of liver cancer tissues and its molecular origin. , 2015, Nanoscale.

[31]  R. Jain,et al.  Antiangiogenesis strategies revisited: from starving tumors to alleviating hypoxia. , 2014, Cancer cell.

[32]  Triantafyllos Stylianopoulos,et al.  The role of mechanical forces in tumor growth and therapy. , 2014, Annual review of biomedical engineering.

[33]  G. Semenza,et al.  Oxygen sensing, hypoxia-inducible factors, and disease pathophysiology. , 2014, Annual review of pathology.

[34]  Derek S. Chan,et al.  Targeting CXCL12 from FAP-expressing carcinoma-associated fibroblasts synergizes with anti–PD-L1 immunotherapy in pancreatic cancer , 2013, Proceedings of the National Academy of Sciences.

[35]  Triantafyllos Stylianopoulos,et al.  Combining two strategies to improve perfusion and drug delivery in solid tumors , 2013, Proceedings of the National Academy of Sciences.

[36]  Rakesh K. Jain,et al.  Angiotensin inhibition enhances drug delivery and potentiates chemotherapy by decompressing tumour blood vessels , 2013, Nature Communications.

[37]  G. Coukos,et al.  Deciphering and reversing tumor immune suppression. , 2013, Immunity.

[38]  Matija Snuderl,et al.  Coevolution of solid stress and interstitial fluid pressure in tumors during progression: implications for vascular collapse. , 2013, Cancer research.

[39]  R. Jain.  Normalizing tumor microenvironment to treat cancer: bench to bedside to biomarkers. , 2013, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[40]  Ueli Aebi,et al.  The nanomechanical signature of breast cancer. , 2012, Nature nanotechnology.

[41]  R. Jain,et al.  Vascular normalizing doses of antiangiogenic treatment reprogram the immunosuppressive tumor microenvironment and enhance immunotherapy , 2012, Proceedings of the National Academy of Sciences.

[42]  R. Jain,et al.  TGF-β blockade improves the distribution and efficacy of therapeutics in breast carcinoma by normalizing the tumor stroma , 2012, Proceedings of the National Academy of Sciences.

[43]  Triantafyllos Stylianopoulos,et al.  Causes, consequences, and remedies for growth-induced solid stress in murine and human tumors , 2012, Proceedings of the National Academy of Sciences.

[44]  R. Jain,et al.  Normalization of tumour blood vessels improves the delivery of nanomedicines in a size-dependent manner , 2012, Nature nanotechnology.

[45]  Pierre Validire,et al.  Matrix architecture defines the preferential localization and migration of T cells into the stroma of human lung tumors. , 2012, The Journal of clinical investigation.

[46]  I. Melero,et al.  Molecular Pathways: Hypoxia Response in Immune Cells Fighting or Promoting Cancer , 2011, Clinical Cancer Research.

[47]  Lin Zhang,et al.  Tumour hypoxia promotes tolerance and angiogenesis via CCL28 and Treg cells , 2011, Nature.

[48]  W. Wilson,et al.  Targeting hypoxia in cancer therapy , 2011, Nature Reviews Cancer.

[49]  J. Schlom,et al.  Tumor-infiltrating immune cells and prognosis: the potential link between conventional cancer therapy and immunity , 2011, Experimental biology and medicine.

[50]  R. Jain,et al.  Losartan inhibits collagen I synthesis and improves the distribution and efficacy of nanotherapeutics in tumors , 2011, Proceedings of the National Academy of Sciences.

[51]  R. Jain,et al.  Delivering nanomedicine to solid tumors , 2010, Nature Reviews Clinical Oncology.

[52]  T. Padhya,et al.  HIF-1α regulates function and differentiation of myeloid-derived suppressor cells in the tumor microenvironment , 2010, The Journal of experimental medicine.

[53]  G. Prud’homme,et al.  Tranilast inhibits the growth and metastasis of mammary carcinoma , 2009, Anti-cancer drugs.

[54]  R. Jain,et al.  Matrix metalloproteinases-1 and -8 improve the distribution and efficacy of an oncolytic virus. , 2007, Cancer research.

[55]  D. Meijer,et al.  The Antiproliferative Drug Doxorubicin Inhibits Liver Fibrosis in Bile Duct-Ligated Rats and Can Be Selectively Delivered to Hepatic Stellate Cells in Vivo , 2006, Journal of Pharmacology and Experimental Therapeutics.

[56]  R. Jain.  Normalization of Tumor Vasculature: An Emerging Concept in Antiangiogenic Therapy , 2005, Science.

[57]  E. Uchida,et al.  Anti-tumor effect of N-[3,4-dimethoxycinnamoyl]-anthranilic acid (tranilast) on experimental pancreatic cancer. , 2002, Journal of Nippon Medical School = Nippon Ika Daigaku zasshi.

[58]  Rakesh K. Jain,et al.  Normalizing tumor vasculature with anti-angiogenic therapy: A new paradigm for combination therapy , 2001, Nature Medicine.

[59]  R. Jain,et al.  Role of extracellular matrix assembly in interstitial transport in solid tumors. , 2000, Cancer research.

[60]  J. Levick.  Flow through interstitium and other fibrous matrices. , 1987, Quarterly journal of experimental physiology.

[61]  R. Reed,et al.  Interstitial fluid pressure in rats measured with a modified wick technique. , 1977, Microvascular research.

[62]  A. Nebreda,et al.  TGF ‐ beta drives immune evasion in genetically reconstituted colon cancer metastasis , 2018 .

[63]  Andreas Stylianou,et al.  Exploring the Nano-Surface of Collagenous and Other Fibrotic Tissues with AFM. , 2017, Methods in molecular biology.

[64]  Rakesh K. Jain,et al.  Taming vessels to treat cancer. , 2008, Scientific American.

[65]  M. Weller,et al.  N-[3,4-dimethoxycinnamoyl]-anthranilic acid (tranilast) suppresses microglial inducible nitric oxide synthase (iNOS) expression and activity induced by interferon-gamma (IFN-gamma). , 2001, British journal of pharmacology.