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Peritoneal Surface Malignancies

P46: TWIST1 as a potential master transcription factor regulating colorectal cancer metastasis

    Author(s)

  • AL

    Alessandro La Ferlita, PhD

    Post Doctoral Scholar
    Department of Cancer Biology and Genetics, The James Comprehensive Cancer Center, The Ohio State University, United States

    • Submitter(s)

  • MP

    Manoj H. Palavalli, MD

    Research Fellow
    Division of Surgical Oncology, The James Comprehensive Cancer Center, The Ohio State University
    Columbus, Ohio, United States

    • Author(s)

  • MP

    Manoj H. Palavalli, MD

    Research Fellow
    Division of Surgical Oncology, The James Comprehensive Cancer Center, The Ohio State University
    Columbus, Ohio, United States

  • MI

    Mozaffarul Islam, PhD

    Senior Research Scientist
    Division of Surgical Oncology, The James Comprehensive Cancer Center, The Ohio State University, United States

  • JW

    Jing Wen, PhD

    Research Scientist
    Division of Surgical Oncology, The James Comprehensive Cancer Center, The Ohio State University, United States

    • ePoster Abstract Author(s)

  • Alex C. Kim, MD, PhD (he/him/his)

    Surgical Oncologist
    UT Southwestern Medical Center
    Dallas, Texas, United States

Introduction: Colorectal cancer peritoneal metastasis (CRC PM) results in poor prognosis with a median overall survival of 16.3 months and almost no survival at 5 years. The poor outcome in patients with CRC PM is a result of recurrent treatment resistance that exhausts therapeutic options. Remarkably, the transcription regulation that triggers CRC PM initiation and progression remains largely unknown. CRC PM is classified as consensus molecular subtype 4 (CMS) which is enriched in genes responsible for self-renewal, invasion, therapeutic resistance, and metastasis.  We identified TWIST1 as a possible regulator of these processes leading to CRC PM.

Methods: RNA-Seq data of liver and peritoneal metastases were retrieved from GEO (GSE50760; GSE183202) and analyzed to assess gene expression. Western blot analysis of patient samples was conducted for transcription factor expression. Metastatic human and mouse cell lines LoVo and CT-26, respectively, were used for shRNA-knockdown and CRISPR-knockout of TWIST1 for in vitro and in vivo experiments. RNA-Seq experiments were performed using LoVo cell line knockdown for TWIST1. Finally, ChIP-Seq with the immunoprecipitation of TWIST1 was performed in LoVo.

Results: Our study revealed that TWIST1 is significantly upregulated in patients with CRC PM.  shRNA and CRISPR knockout of TWIST1 revealed a significant decrease in invasion and migration in human (LoVo) and mouse (CT-26) colorectal cancer cell lines. RNA-Seq analysis of TWIST1 knockdown demonstrated significant downregulation of key genes in pathways involved in EMT (i.e., TGF-β, SMURF2), extracellular matrix production and regulation (MMP25, MMP19), mucus regulation (MUC16, MUC4, GLI1), cell motility (MSN), and self-renewal (CD44). Furthermore, ChIP-Seq analysis showed direct binding to key genes in the aforementioned pathways. Finally, syngeneic mice with CRISPR-mediated TWIST1 knockout in CT-26 cells demonstrated improved survival and decreased tumorigenesis.

Conclusions: TWIST1 is a potential master transcription factor critically involved in tumorigenesis and chemoresistance in CRC PM.

Learning Objectives:

  • Upon completion, participant will be able to describe the role of TWIST1 in CRC PM.
  • Upon completion, participant will be able to recognize important genes responsible for CRC PM tumorigenesis and chemoresistance.
  • Upon completion, participant will be able to discover novel targets for CRC PM