# Three-dimensional organoid models to study breast cancer progression

> **NIH NIH R37** · UNIVERSITY OF PITTSBURGH AT PITTSBURGH · 2022 · $418,813

## Abstract

Approximately 20% of breast cancers detected through mammography are pre-invasive Ductal Carcinoma in
situ (DCIS). If left untreated, approximately 20-50% of DCIS will progress to more deadly Invasive Ductal
Carcinoma (IDC). No prognostic biomarkers can reliably predict the risk of progression from DCIS to IDC. Similar
genomic profiles of matched pre-invasive DCIS and IDC suggests that the progression is not driven by genetic
aberrations in DCIS cells, but microenvironmental factors, such as hypoxia and metabolic stress prevalent in
DCIS, may drive the transition. We need innovative models to investigate how to halt steps of DCIS progression
to invasive phenotypes and subsequent metastasis from the primary site. This proposal directly addresses
this unmet need by developing a novel three-dimensional in vitro organoid model that recapitulates key
hallmarks of DCIS to IDC progression: tumor-size induced hypoxia and metabolic stress, tumor heterogeneity
and spontaneous emergence of migratory phenotype in the same parent cells without any additional stimulus. A
tangible advantage of the proposed organoid models is the ability to precisely and reproducibly study how the
hypoxic microenvironment induces tumor migration in real time and in isolation from non-tumor cells present in
vivo, providing unique opportunity to define tumor-intrinsic mechanisms of DCIS to IDC progression. Our
preliminary observations lead to central hypothesis that tumor size-induced hypoxia establishes a “hypoxic
secretome”, which initiates the migratory phenotype; the hypoxic secretome then cooperate with intracellular
signaling networks to independently maintain cell migration. We propose three independent but inter-related
aims to link hypoxic secretome with the initiation, maintenance and spatial distribution of migratory phenotypes.
Aim 1 will engineer size-controlled DCIS organoids (150-600 µm) with controlled hypoxic microenvironments to
identify and examine how hypoxic secretome initiates migratory phenotype. We will combine experimental
organoid models with time-lapse imaging and computational approaches to study organoid migration. Aim 2 will
demonstrate that migratory cells can re-establish the secretome and maintain migratory phenotype independent
of hypoxia. We will reconstruct an intracellular signaling network activated by the hypoxic secretome using
microarray data. We will verify these gene expression signatures in sorted migratory and non-migratory cells,
and validate them using secretome inhibition studies. Aim 3 will investigate, for the first time, the spatial
distribution and origin of the migratory phenotype. We will use CRISPR-based gene knock-in (FP-labeling),
automated image analyses, and a deep-learning algorithm to track and visualize the emergence of migratory
phenotypes from the hypoxic core outward to the periphery or from the migratory front.
The successful development of this 3D organoid model and completion of the proposed work will provide
answ...

## Key facts

- **NIH application ID:** 10438709
- **Project number:** 5R37CA232209-05
- **Recipient organization:** UNIVERSITY OF PITTSBURGH AT PITTSBURGH
- **Principal Investigator:** Shilpa Sant
- **Activity code:** R37 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $418,813
- **Award type:** 5
- **Project period:** 2018-07-01 → 2023-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10438709

## Citation

> US National Institutes of Health, RePORTER application 10438709, Three-dimensional organoid models to study breast cancer progression (5R37CA232209-05). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10438709. Licensed CC0.

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