# Structure-based engineering of immune cytokine signaling

> **NIH NIH R01** · STANFORD UNIVERSITY · 2023 · $517,818

## Abstract

ABSTRACT:
Immunoregulatory cytokines engage transmembrane signaling receptors in order to mediate a wide range of
functions including leukocyte proliferation, differentiation, and expansion through JAK/STAT activation. Most
immunoregulatory cytokines possess both redundant and distinct activities that are critical to normal immune
homeostasis, but this functional pleiotropy presents a major problem for the effective use of these cytokines as
immunotherapeutic cancer drugs. Cytokine pleiotropy is a consequence of different cytokine receptors being
expressed on multiple different counterbalancing cell types that serves to neutralize anti-tumor actions and lead
to systemic toxicity. During the prior term of this award, we gained an appreciation for the extracellular
structural architectures of a spectrum of different immune cytokine complexes, including those of IL-2, IL-10,
IL-12, IL-21, IL-22, IL-23, IL-27, IFN, and IFN. In this renewal application, with these structural templates in
hand, in Aim 1 we focus our studies on the cytokines IL-2, IFN, and IL-12. These immune master regulators
engage different but overlapping branches of the immune system, and share the issues of pleiotropy and toxicity
that, if uncoupled, could lead to powerful cancer immunotherapeutic agents. We propose to “tune” signaling
through structure-based cytokine engineering to attempt to create variants with decoupled pleiotropy, cell subset
preferences, enhanced anti-tumor efficacies, and reduced toxicity – both alone and in combination. In Aim 2, we
wish to understand the mechanistic basis for how tuned cytokines can differentially activate signaling inside the
cells. Based on a recent breakthrough in our 20-year quest to solve the full-length JAK structure, we continue to
pursue structural information on how cytokine binding to their receptors activates Janus Kinase (JAK)
molecules, by reconstituting and imaging activated JAK homo- and heterodimers bound to both cytokine and
intracellular JAKs and STATs. In this fashion, by combining structural biology, protein engineering, cell
signaling, and in vivo tumor studies, we propose to obtain a complete molecular snapshot of cytokine receptor
signaling from the initial engagement of ligand through the activation of intracellular signaling cascades and
leverage this information for the engineering of cancer immunotherapeutics.

## Key facts

- **NIH application ID:** 10717246
- **Project number:** 2R01AI051321-21
- **Recipient organization:** STANFORD UNIVERSITY
- **Principal Investigator:** Kenan Christopher GARCIA
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $517,818
- **Award type:** 2
- **Project period:** 2002-04-01 → 2028-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10717246

## Citation

> US National Institutes of Health, RePORTER application 10717246, Structure-based engineering of immune cytokine signaling (2R01AI051321-21). Retrieved via AI Analytics 2026-06-11 from https://api.ai-analytics.org/grant/nih/10717246. Licensed CC0.

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