Search

Transforming growth factor beta (TGF-β) isoforms are the primary mediators for TGF-β signaling via TGF-β receptors and downstream phosphorylation/dephosphorylation cascade. TGF-β is associated with a wide range of biological processes in oncology, including tumor cell invasion, migration, angiogenesis, immunosuppression, as well as regulation of tumor stem cell properties. Mouse 4T1 mammary carcinoma cell line is a transplantable tumor cell line that is highly tumorigenic and invasive and, unlike most tumor models, can spontaneously metastasize from the primary tumor in the mammary gland to multiple distant sites including lymph nodes, blood, liver, lung, brain, and bone. Considering rather challenging preclinical evaluation of antitumor activity in tumor models, mouse 4T1 mammary carcinoma model has been widely used in the literature for evaluation of TGF-β antagonists. In this report, we describe the efficacy of ISTH0047 - a potent and selective TGF-β2 antisense oligonucleotide - in murine 4T1 primary tumors and lung metastasis following tumor cell injection into the mammary fat pad (orthotopic tumor model) of syngeneic Balb/c mice. Consistent with literature data generated with other classes of TGF-β antagonists (e.g., small-molecule kinase inhibitors, antibodies or TGF-β trap agents), although limited antitumor activity was demonstrated on primary tumor growth, marked and statistically significant decrease of lung metastasis number was observed upon subcutaneous administrations of ISTH0047. In addition, side by side comparison with murine surrogates of CTLA-4 or PD-1 antibodies indicated similar efficacy of all test items on lung metastasis in this model. Taken together, these encouraging results pave the way for in-depth preclinical evaluation of both ‘seed and soil’ theory and efficacy of combination regimen (immunomodulation) for better clinical outcome. Citation Format: Katja Wosikowski, Kathy Hasenbach, Jutta Petschenka, Diana Barea Roldan, Sebastian Kreiter, Ugur Sahin, Guillaume Serin, Julie-Orlane Redon, Marc Hillairet de Boisferon, Francis Bichat, Hanna Kohonen, Michel Janicot. Antimetastatic activity of ISTH0047, a potent and selective TGF-beta 2 antisense oligonucleotide, in syngeneic lung metastatic model of mouse 4T1 mammary carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3741.

Transforming growth factor beta (TGF-β) is a key member of a large family of cytokines, which play critical, pleiotropic roles in the pathophysiology of various human diseases, such as cancer, inflammation, autoimmune disease, and cirrhosis/fibrosis. TGF-β1, -2 and -3 isoforms are cytokines encoded by different genes but sharing strong sequence and structure homology. They function as the primary mediators of TGF-β signaling via both non-canonical and canonical signaling pathways. In Oncology, TGF-β isoforms are associated with a wide range of biological processes such as tumor cell invasion and migration, angiogenesis, immunosuppression, as well as regulation of tumor stem cell properties. Hence, blocking the TGF-β signaling pathway may have a multifold therapeutic benefit in Oncology, although therapeutic relevance of the respective TGF-β isoforms remains poorly documented. In order to evaluate the specific biological relevance of TGF-β1 isoform in cancer, we have initiated an extensive discovery program for identification of antisense oligodeoxynucleotide constructs selectively inhibiting expression of the TGF-β1 ligand. Based on the sequence of the human TGF-β1 mRNA, more than 150 Locked Nucleic Acid (LNA)-modified gapmers were designed, synthesized and tested in cell-based assays. Highly potent and selective TGF-β1 constructs were identified and selected based on efficient suppression of TGF-β1 mRNA/protein expression in different human and rodent tumor cell lines, and in human Peripheral Blood Mononuclear Cells (PBMCs). Effective target downregulation was demonstrated after lipofectamine aided-transfection (sub-nM concentration range), but also in the absence of any transfection agent (gymnotic delivery) at sub-µM concentrations. Selected TGF-β1 specific oligonucleotides were further tested in relevant animal xenograft models and strong target inhibition was also observed following systemic administration. Further details of this discovery program will be discussed and pharmacology properties and features of potent and selective TGF-β1 antagonists will be presented. Citation Format: Frank Jaschinski, Hanna Korhonen, Stephan Braun, Katja Wosikowski, Michel Janicot. Novel potent antisense oligonucleotides targeting transforming growth factor beta1 (TGF-β1). [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 717. doi:10.1158/1538-7445.AM2014-717

Transforming growth factor beta (TGF-β represents a family of cytokines, which function as the primary mediators for TGF-β signaling via TGF-β receptor type II (TβIIR) and both non-canonical and canonical downstream signaling pathways. TGF-β is associated with a wide range of biological processes in oncology, including tumor cell invasion, migration, angiogenesis, immunosuppression, as well as regulation of tumor stem cell properties. Hence, optimal preclinical evaluation of efficacy of TGF-β antagonists is challenging. Isarna Therapeutics has designed and developed selective and potent LNA-modified antisense oligonucleotides targeting the various TGF-β isoforms. In order to adequately evaluate selected preclinical development candidates, Oncodesign has developed customized experimental Renca tumor models in syngeneic and/or immunodeficient mice. The Renca cell line was established from a murine transplantable renal adenocarcinoma of spontaneous origin, and has been used under various experimental conditions: (1) subcutaneous tumor model by inoculating cells into the flanks of the animals; (2) the pulmonary metastatic tumor model by an intravenous injection of cells into the tail vein; and (3) the orthotopic tumor model by injecting cells into the renal subcapsule (and subsequent pulmonary metastasis). Outcome of this development program and preliminary results for selected TGF-β antisense oligonucleotides will be presented and discussed. Citation Format: Hanna Korhonen, Julie-Orlane Redon, Damien France, Guillaume Serin, Francis Bichat, Frank Jaschinski, Katja Wosikowski, Michel Janicot. Renca RCC syngeneic model to evaluate efficacy of novel antisense oligonucleotides targeting TGF-β isoforms. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3126. doi:10.1158/1538-7445.AM2014-3126

Transforming growth factor beta (TGF-β) is a key member of a large family of cytokines, including bone morphogenetic protein (BMP), nodals, activins and others, which play critical, pleiotropic roles in the pathophysiology of various human diseases, such as cancer, inflammation, autoimmune disease, and cirrhosis/fibrosis. In particular, the different TGF-β isoforms (TGF-β1, -β2, and -β3, encoded by different genes but sharing high sequence and structure homology) are overexpressed in many human tumors. Correlations between TGF-β expression, cancer stage and clinical parameters have been reported and linked to poor clinical outcome. TGF-β has been associated with a wide range of tumor-promoting processes, including tumor cell invasion and migration, angiogenesis, immunosuppression, as well as tumor stem cell maintenance and protection. Therefore, blocking the TGF-β signaling pathway via inhibition of TGF-β expression appears as an attractive therapeutic intervention in Oncology. We have previously reported the rational design and preliminary outcome of an extensive discovery program for identification of antisense oligodeoxynucleotides targeting the various TGF-β isoforms. We present here ASPH_0047, a 17-mer LNA-modified gapmer based on the sequence of the human TGF-β2 mRNA, as novel preclinical development candidate. ASPH_0047 shows potent and selective target downregulation (mRNA and protein) in various cell-based assays. Preliminary preclinical biodistribution studies in rodent point at tissue penetration in expected target organs (e.g., liver, kidney, spleen) and accumulation in tumor tissue consistent with observed target downregulation and efficacy in relevant in vivo experimental antitumor models. Key pharmacology properties and preclinical features of ASPH_0047 supporting rapid advancement to clinical development will be presented. Citation Format: Katja Wosikowski, Frank Jaschinski, Hanna Kohonen, Stephan Braun, Eugen Leo, Michel Janicot. ASPH_0047: a potent and selective antisense oligonucleotide-targeting transforming growth factor beta 2 (TGF-β2). [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 716. doi:10.1158/1538-7445.AM2014-716