# Live cell reporters of genetic changes in stiff vs soft surroundings - Causes & Consequences

> **NIH NIH U01** · UNIVERSITY OF PENNSYLVANIA · 2022 · $66,580

## Abstract

Project Summary/Abstract
Aneuploidy is defined as the existence of an abnormal number of chromosomes in a cell, and it has long been a
defining characteristic of cancers. 90% of all solid tumors, for example, show chromosomal gains/losses. Gaining
and/or losing chromosomes generates phenotypic heterogeneity within tumors that can be beneficial to survival
by creating subpopulations that may become resistant to therapeutics, providing new mechanisms to evade
immune cells, facilitating growth and proliferation in challenging microenvironments, among other advantages.
However, we currently struggle to easily identify aneuploid cells—typically we need to kill them for sequencing,
meaning that we can only at best infer possible causes, consequences, and possible long-term viability and
evolution. This also limits our ability to identify how aneuploid cells interact with other cells and study if and how
these interactions differ from other “less” aneuploid cells. These interactions become increasingly important
when the engaging cell is from the immune repertoire.
 In this proposed research, we tackle two aspects of aneuploidy: (1) providing a simple, effective method
to easily identify aneuploidy in live cells and (2) understanding how aneuploidy can modulate macrophage-
mediated phagocytosis of cancer cells. We already have successfully generated several chromosome reporters,
in which we fuse fluorescent proteins to only single alleles of constitutively expressed genes. For these
appropriate genes, we find that fluorescence loss equates to chromosomal loss—thoroughly characterized from
genomic PCR to deep single-cell sequencing. We further seek to expand this toolkit to make it accessible to
more appropriate mouse models. On the immunobiology end, we already find that high levels of aneuploid help
promote an initial macrophage-immune response. This suggests that initial stages of aneuploidy (before a cancer
can adapt and evolve to tolerate it) are susceptible to macrophage clearance. Although seemingly two distinct
ideas, we finally plan to merge our reporter approach in analyzing how macrophages are phenotypically molded
by aneuploid cells that we can easily identify. Do macrophages eat/clear these reporter-negative aneuploid cells?
Are reporter-negative cells simply more susceptible to macrophages or are profound paracrine signaling
pathways involved? We will extend our studies to both in vitro and in vivo models to better characterize
macrophage response under these circumstances, and we will also ultimately combine these studies with
powerful single-cell sequencing to see how aneuploidy sculpts macrophage behavior and their overall effect on
the immune landscape.

## Key facts

- **NIH application ID:** 10594852
- **Project number:** 3U01CA254886-02S1
- **Recipient organization:** UNIVERSITY OF PENNSYLVANIA
- **Principal Investigator:** Dennis E. Discher
- **Activity code:** U01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $66,580
- **Award type:** 3
- **Project period:** 2021-04-01 → 2026-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10594852

## Citation

> US National Institutes of Health, RePORTER application 10594852, Live cell reporters of genetic changes in stiff vs soft surroundings - Causes & Consequences (3U01CA254886-02S1). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10594852. Licensed CC0.

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