# Immunotherapy Modeling in Organoids Co-preserving Tumor and Infiltrating Immune Compartments

> **NIH NIH R01** · STANFORD UNIVERSITY · 2024 · $623,480

## Abstract

PROJECT SUMMARY
The immune system remarkably distinguishes between self and non-self/self-aberrant antigens, affording
exquisite anti-tumor specificity and inhibition of tumorigenesis. However, tumor immunosurveillance is
unfortunately opposed by tumor cell evasion of the immune response. Immune checkpoint blockade (ICB)
targeting PD-1, PD-L1, CD40 and others, as well as adoptive cell transfer (CAR-T, bulk TILs) favorably modulate
this equilibrium for therapeutic benefit. However, response rates are often incomplete, progressive disease is
common, and predictive biomarkers are suboptimal.
 The development of next-generation immunotherapies has been hindered by a lack of in vitro models
that functionally recapitulate syngeneic interactions between tumor and infiltrating immune cells. In response,
we have developed organoid methods that culture primary human tumor biopsies together with their infiltrating
immune components as a cohesive unit without reconstitution. These “patient-derived tumor organoids” (PDO)
preserve tumor cells alongside endogenous T, B, NK cells and macrophages, robustly recapitulate the T cell
receptor clonotype repertoire of the original tumor, and crucially, manifest tumor-infiltrating lymphocyte (TIL)
expansion, activation and tumor cell killing in response to anti-PD-1/PD-L1 therapeutic antibodies (Cell, 2018).
The PDO system thus represents a holistic organoid model of human tumor-immune interactions.
 Here, we leverage the PDO technique to investigate immunotherapeutic mechanisms and
treatments in PD-1-responsive cutaneous squamous cell carcinoma (cSCC) and melanoma, exploiting
pre- and post-treatment human biopsies and mouse models. Aim 1 hypothesizes that checkpoint inhibition
induces a complex and sequential network response involving immune-tumor and immune-immune crosstalk.
Thus, Aim 1 employs the ability to perform serial time-course sampling of PDOs to define a single cell RNA-seq
network cellular crosstalk model of the early anti-PD-1-stimulated anti-tumor immune response over multiple
acute time points typically inaccessible to clinical biopsies performed after months. Importantly, comparison of
this immune propagation in responding versus non-responding mouse and human organoids will define nodal
points conferring resistance. Aim 2 improves bulk TIL adoptive transfer immunotherapy by using PDOs as living
bioreactors to enrich tumor-reactive mouse and human melanoma TILs by anti-PD-1 checkpoint inhibition,
followed by testing of enhanced anti-tumor activity in vitro and in vivo. Lastly, Aim 3 performs a co-treatment
trial comparing anti-PD-1 responses of pre-treatment biopsy cSCC PDOs to clinical outcomes. Further, post-
treatment biopsy PDOs are re-challenged with anti-PD-1 and a novel agent inactivating PD-1 by
dephosphorylation. We thus utilize the holistic PDO model preserving endogenous tumor epithelial and immune
components en bloc to investigate and improve cancer immunotherapy via our team of Calvin Kuo ...

## Key facts

- **NIH application ID:** 10795111
- **Project number:** 5R01CA251514-04
- **Recipient organization:** STANFORD UNIVERSITY
- **Principal Investigator:** CALVIN J KUO
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $623,480
- **Award type:** 5
- **Project period:** 2021-03-17 → 2026-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10795111

## Citation

> US National Institutes of Health, RePORTER application 10795111, Immunotherapy Modeling in Organoids Co-preserving Tumor and Infiltrating Immune Compartments (5R01CA251514-04). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10795111. Licensed CC0.

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