# Architectural regulation of cytotoxic synapse detachment

> **NIH NIH R21** · SLOAN-KETTERING INST CAN RESEARCH · 2022 · $265,500

## Abstract

Summary
Cytotoxic lymphocytes, comprising cytotoxic T lymphocytes (CTLs) and natural killer cells, kill by forming
specialized immune synapses with their targets, into which they channel toxic factors that induce apoptosis.
Although much is known about how cytotoxic synapses form, the cellular and molecular mechanisms that control
their dissolution are poorly understood. Addressing this gap in knowledge is important because cytotoxic
lymphocytes must let go of dying cells to kill multiple targets in a serial manner. Efficient synapse disassembly
also prevents spurious inflammation by attenuating sustained cytotoxic lymphocyte activation and also by
facilitating the clearance of apoptotic corpses by phagocytes. Prior attempts to study this process have focused
on the recognition of biochemical features associated with cell death. Our preliminary studies, however, suggest
an alternative and conceptually innovative model in which lymphocyte detachment is induced by biophysical
changes in dying target cells. We are particularly interested apoptotic contraction, which we have found occurs
just before the dissolution of CTL-target cell conjugates. In addition, genetic and pharmacological decoupling of
contraction from apoptosis delays the dissociation response. Building upon these findings, we hypothesize that
CTLs use the characteristic biophysical features of contracting targets to trigger release. Our proposed studies,
which are divided into two Specific Aims, will explore the biophysical basis for CTL dissociation and the molecular
pathways that govern the process. Specific Aim 1 will employ single cell mechanobiological methods to assess
the effects of apoptotic contraction on cortical rigidity and surface ligand mobility. We will also use an
optogenetics approach to determine whether cytoskeletal contraction is sufficient to induce synapse
disassembly. Specific Aim 2 will examine the molecular bases for apoptotic contraction and its detection by
CTLs, using targeted loss-of-function and CRISPR/Cas9 screening. Our proposed studies will leverage
technically innovative methods, including super-resolution imaging, optogenetics, and atomic force microscopy.
They will also introduce a novel concept, that mechanosensing can determine the lifetime of an immune cell-cell
interaction. The successful completion of this project will address a long-standing enigma in lymphocyte cell
biology and likely reveal new avenues for the modulation of cellular cytotoxicity in the clinic. As such, this
proposal is highly relevant to the NIH mission in that it will contribute to the advancement of knowledge that could
improve human health.

## Key facts

- **NIH application ID:** 10467438
- **Project number:** 1R21AI169847-01
- **Recipient organization:** SLOAN-KETTERING INST CAN RESEARCH
- **Principal Investigator:** Morgan A Huse
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $265,500
- **Award type:** 1
- **Project period:** 2022-03-01 → 2024-02-29

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10467438

## Citation

> US National Institutes of Health, RePORTER application 10467438, Architectural regulation of cytotoxic synapse detachment (1R21AI169847-01). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10467438. Licensed CC0.

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