# Engineering CARs against cancer-specific glycoproteins

> **NIH NIH R21** · UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN · 2020 · $208,452

## Abstract

Abstract
Adoptively transferred T cells expressing chimeric antigen receptors (CARs) have shown success in
eliminating hematopoietic cancers, but have so far been challenging in the solid tumor setting. A related
challenge has been the identification of targets that will not be lost upon selective pressures exerted by the
treatment (i.e. antigen loss variants). In this project, we propose to use protein engineering of the antigen
binding domains of an antibody against novel targets that will optimize the therapy yet not be subject to antigen
loss. The targets are cancer cell surface glycoproteins that contain a single, O-linked GalNAc-linkage, called a
Tn antigen. These Tn-antigens are not found on the surface of normal cells, but have long been known to be
associated with cancer growth and metastasis. Over the past few decades, three particular targets of Tn-linked
expression, MUC1, MUC4, and MUC16 (CA125) have been described, especially as antigens on various solid
tumors including ovarian cancers. In this project, we will use yeast display to engineer the Tn-dependent
antibody called 237 that we have already shown can be formatted as CAR T cells, yet which can be improved
for both potency and range of targetable cancers. The project will involve multidisciplinary expertise, ranging
from protein engineering to cancer immunology. Guided by the known crystal structure of the 237 Fab:Tn-
antigen complex, we will use yeast display and directed evolution to: i) generate 237 scFv with a range of
affinities, and ii) broaden the cancer-specific reactivity to additional Tn-peptide backbones, including MUC1
and MUC4. Our Specific Aims are: Aim 1. To engineer a range of affinities and novel specificities using 237
scFv fragments as a scaffold. Aim 2. To determine the pre-clinical efficacy of engineered anti-GalNAc CARs in
eradicating solid tumors. The results will provide a blueprint for the effective use and development of next-
generation Tn-dependent CARs. Through this proposal, these CARs will be engineered to exhibit a high
degree of specificity and activity for cancers with aberrant glycosylation due to cancer-specific mutation of
Cosmc or dysregulation of glyco-transferases.

## Key facts

- **NIH application ID:** 9968189
- **Project number:** 5R21CA238628-02
- **Recipient organization:** UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
- **Principal Investigator:** David M. Kranz
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $208,452
- **Award type:** 5
- **Project period:** 2019-07-01 → 2022-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9968189

## Citation

> US National Institutes of Health, RePORTER application 9968189, Engineering CARs against cancer-specific glycoproteins (5R21CA238628-02). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/9968189. Licensed CC0.

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