# Understanding the role of TWIST1 in colorectal cancer progression and metastasis

> **NIH NIH F30** · HARVARD MEDICAL SCHOOL · 2024 · $38,135

## Abstract

Project Summary:
Although almost all colorectal cancer (CRC) mortality is due to metastasis, there are currently no drugs to prevent
or halt metastasis. A better understanding of the underlying biology of CRC progression and metastasis could
lead to the next generation of therapeutics aimed at blocking steps in the metastatic cascade. Previous research
provided a framework to understand tumor metastasis through sequential stages of cancer cell invasion into the
basement membrane, migration into vasculature, circulation, extravasation, and colonization of distant organs.
Each step is heavily dependent on diverse cell-cell communications and the microenvironment, which has not
been well recapitulated in previous in vitro and in vivo model systems to study CRC metastasis. However, the
recent development of CRISPR-engineered 3D colorectal cancer organoids (CRCOs) and colonoscopy-guided
orthotopic CRC transplantation models have provided a more physiological model system to study metastasis.
The CRCOs are precisely engineered with oncogenic mutations found in the majority of human CRC, such as
mutations in the Apc, Kras, and P53 genes (AKP). Moreover, these engineered CRCOs are transplanted into
the murine colon, where they grow in their native environment and metastasize to the liver: the most common
site of CRC metastasis in humans. Previous research using these model systems identified Twist1 as
significantly upregulated in CRC liver metastases compared to the primary tumor. Twist1 is a well-known
transcription factor involved in promoting epithelial-mesenchymal transition (EMT). EMT is when epithelial cells
lose their cell adhesions and acquire a more motile mesenchymal phenotype. It is unclear what role Twist1 plays
in CRC progression and metastasis. Aim 1 will verify that Twist1 is upregulated in liver metastases compared to
the primary tumor from AKP CRCOs in orthotopically transplanted mice using immunohistochemistry,
RNAscope, and RT-qPCR. Aim 1 will also assess whether Twist1 is necessary for liver metastasis formation
through CRISPR-Cas9-mediated loss in AKP organoids and a diphtheria toxin receptor cell-elimination model.
Moreover, to evaluate if overexpression of Twist1 increases metastatic potential, the AKP CRCO will be
engineered to overexpress Twist1 using complementary DNA. Thus, Aim 1 will determine whether Twist1 is
required for CRC metastasis. The role of TWIST1 in human CRC progression and metastasis will be the focus
of Aim 2. Aim 2 will screen human CRC tissue microarrays for expression of TWIST1 and other EMT
markers. TWIST1 in each tissue will be quantified and correlated with patient tumor site, stage, genotype, and
outcome. Moreover, to validate TWIST1 is more highly expressed in human CRC metastases, a human CRCO
will be transplanted into immunodeficient mice and assayed for expression of TWIST1 by immunohistochemistry,
RNAscope, and RT-qPCR. Such knowledge of TWIST1’s role in CRC metastasis can illuminate targets for the
...

## Key facts

- **NIH application ID:** 10847429
- **Project number:** 5F30CA260789-04
- **Recipient organization:** HARVARD MEDICAL SCHOOL
- **Principal Investigator:** Joseph Charles Sedlak
- **Activity code:** F30 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $38,135
- **Award type:** 5
- **Project period:** 2021-07-01 → 2024-11-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10847429

## Citation

> US National Institutes of Health, RePORTER application 10847429, Understanding the role of TWIST1 in colorectal cancer progression and metastasis (5F30CA260789-04). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10847429. Licensed CC0.

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