Abstract 3940: Validation of TFX1, a first-in-class target for cancer metastasis

Abstract

Abstract Most of the drugs approved for metastatic epithelial carcinomas treat the proliferation of the tumor and not the biology of metastasis. To date, all clinical trials around metastasis that focused on proteases, kinases, or integrins have failed in clinics. As a result, metastasis biology-focused drug discovery has been deprioritized.We asked the question as to what the rate-limiting steps of metastasis were. To answer it, we created a cell-based functional assay platform, METAssay™, that dissected all metastasis biology steps into thirty cellular assays and characterization steps. The platform identified the functional differences between metastatic and proliferating cells. This platform was normalized with patient tumor cells, and a machine learning algorithm, METSCAN™, was subsequently applied to identify the weighted steps that contributed towards successful metastasis. This led to the identification of four first-in-class targets; two transcription factors, one cytokine receptor and one nuclear hormone receptor.We then validated the relevance of the first target, TFX1, by overexpressing it in both colorectal and triple-negative breast cancer cell lines. Interestingly, apart from the expected increase in migration and invasion, all cell lines showed an increase in ROS, Autophagy, PDL-1, CD-73, quiescence, exosome uptake and a decrease in apoptosis, mitosis, and exosome secretion. Conversely, the downregulation of TFX1 showed the opposite effect. TFX1 was purified, and a biochemical isothermal calorimetry (ITC) assay was standardized with a tool compound with a KD of 107 nM. The compound effectively reduced metastatic properties in the cell lines and in multiple patient samples, as analyzed by the METAssay™ platform. Colorectal cancer cell lines overexpressing TFX1 also showed increased platelet binding, which was reduced by the tool compound, but not triple-negative cancer cell lines. Also, the effect of overexpression in non-metastatic cells was more profound than in metastatic cells, suggesting a saturating effect. Further characterization of the tool compound is currently ongoing to help establish a baseline for a novel discovery programme.The ideal drug to delay metastasis should be given to primary tumor patients with no evidence of clinical metastasis, irrespective of their pathological grading. Therefore, it should have a high bar for safety. Our goal is to collaborate on a discovery journey to identify candidates and create a companion diagnostic that will identify primary tumor patients at high risk of metastasis, thereby selecting the right patient cohort for clinical trials. Citation Format: Arnab Roy Chowdhury, Manoj Pandre, Chinmaya Amarkanth, Debabani Roy Chowdhury, John Ellingboe. Validation of TFX1, a first-in-class target for cancer metastasis. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3940.