# Role of a Two-Factor Genetic Circuit Regulating Stemness in Colorectal Cancer

> **NIH NIH F30** · HARVARD MEDICAL SCHOOL · 2022 · $51,752

## Abstract

Project Summary
 Colorectal cancer (CRC) is the third leading cause of cancer death worldwide. Cancer stem cells
(CSCs) in the colon are the source of tumor initiation, proliferation, chemoresistance, and metastasis,
and are therefore considered to adversely impact patient outcome. However, there are no therapies targeting
CSCs for treatment of CRC. Pluripotency factors have been proposed to drive stemness in CSCs, but the
molecular mechanisms remain unclear. Understanding the molecular regulators that enable stemness and
tumorigenicity in CSCs will identify novel therapeutic targets.
 The role of pluripotency factors is best understood in embryonic stem cells (ESCs), where their interplay
has been proposed to drive heterogeneity. We recently identified a potential transcriptional circuit between
pluripotency factors Klf4 and Zfp281 that we hypothesize enables ESCs to switch between specific stem cell
states and thus generate transcriptional heterogeneity. Notably, in CRC, Klf4 and Zfp281 have also implicated
as a tumor suppressor and pro-proliferative agent, respectively. Klf4 inhibits proliferation and expression of
Bmi1, an epigenetic marker that we previously identified as required for CRC tumorigenesis, whereas
Zfp281 is often co-expressed with Bmi1. Klf4 and Zfp281 display reciprocal expression in both ESCs and CRC
cells, but their correlation with stemness is reversed in the two contexts, such that Klf4 is associated with more
stem-like ESCs but more differentiated CRC cells. The role of pluripotency factor circuits has not been
addressed in CSCs, and the precise interplay between Klf4 and Zfp281 is unknown in either ESCs or CRCs. I
hypothesize that during both embryonic and cancer development, Klf4 and Zfp281 oppose each other,
forming a genetic circuit that modulates the balance between stemness and differentiation. I further
propose that perturbing this genetic circuit will restrict stemness and tumorigenicity.
 In this proposal, I will investigate how Klf4 and Zfp281 interact and regulate stemness in embryonic and
tumor development. Aim 1 will define the interactions of Klf4 and Zfp281 that generate opposing stem cell
states in cultured ESCs. Aims 2 and 3 will examine the role, and consequences of perturbation, of Klf4 and
Zfp281 in regulating stemness and tumorgenicity of CRC cells. Specifically, Aim 2 will use CRC lines and
organoids to determine whether Klf4 and Zfp281 knockout restricts the differentiation capacity and expression
of stemness programs in CSCs. Aim 3 will establish the interplay of Klf4 and Zfp281 with each other, and also
Bmi1, in CRC and determine how these contribute to tumorgenicity in vivo. This project will establish the
role of Klf4 and Zfp281 in stemness and tumorigenicity and the nature of the circuit between them. The
molecular and mechanistic insights of this project will further our understanding of how CSCs initiate and grow
tumors, as well as identify novel therapeutic targets for CRC treatment.

## Key facts

- **NIH application ID:** 10364645
- **Project number:** 5F30CA260739-02
- **Recipient organization:** HARVARD MEDICAL SCHOOL
- **Principal Investigator:** Sofia Hu
- **Activity code:** F30 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $51,752
- **Award type:** 5
- **Project period:** 2021-06-01 → 2025-10-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10364645

## Citation

> US National Institutes of Health, RePORTER application 10364645, Role of a Two-Factor Genetic Circuit Regulating Stemness in Colorectal Cancer (5F30CA260739-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10364645. Licensed CC0.

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