# Identifying Resident and Immune Cell Injury Responses in the Human Kidney

> **NIH NIH R21** · YALE UNIVERSITY · 2020 · $251,250

## Abstract

Kidney function, both under normal conditions and in the setting of injury or disease, is predominantly
controlled by the amount, localization and activation state of proteins in specific cells at specific sites and under
specific spatial influences that in turn regulate cell differentiation, division, metabolism, morphology, membrane
polarization, secretion and transport. Imaging mass cytometry (IMC) takes advantage of laser ionization and
time of flight mass spectrometry to simultaneously identify >40 metal ion-conjugated antibodies from tissue
sections at 1micron resolution with essentially no background signal. By mixing cell specific antibodies and
activation state-specific antibodies in the same cocktail, IMC produces a spatially preserved analysis of the
location and activation state of multiple cell types from the same tissue section. We have developed a panel of
21 antibodies for simultaneous kidney section hybridization and IMC analysis, and a machine-learning based
analysis pipeline that has allowed us to provide the first quantitative atlas of the number and location of the
tubular, endothelial, glomerular and interstitial cells in the normal human kidney (Singh et al., JCI-Insight, in
press). We now propose to validate an additional 22-29 antibodies for an expansion of this panel so as to
provide information on the immune activation state and resident epithelial and endothelial cell injury, survival
and death pathway responses predicted to be important in the injured human kidney. We will first perform in
vitro validation of antibodies that detect cell injury/dedifferentiation states, survival pathways (autophagy,
unfolded protein response, proliferation), and cell death/senescence pathways (apoptosis, necroptosis,
pyroptosis, senescence, Specific Aim 1), as well as immune cell populations and their activation states
(Specific Aim 2). We will then perform IMC/Kidney-MAPPS based validation of the composite panel using
carefully characterized human kidney biopsies from patients with defined clinical insults and morphologically
quantified tubular and endothelial injury (Specific Aim 3). The end product of this proposal will be to provide a
validated set of antibodies that can be used to accurately quantify cell injury states and subsequent responses
across a spectrum of human kidney diseases. These antibodies can then be used by investigators who
perform multiplexed imaging of the kidney, including IMC, IONpath MIBI cytometry, Tissue Cytometry and
Repeated Fluorescent Imaging to mechanistically interrogate human AKI.

## Key facts

- **NIH application ID:** 9954680
- **Project number:** 1R21AG067335-01
- **Recipient organization:** YALE UNIVERSITY
- **Principal Investigator:** LLOYD G CANTLEY
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $251,250
- **Award type:** 1
- **Project period:** 2020-05-01 → 2022-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9954680

## Citation

> US National Institutes of Health, RePORTER application 9954680, Identifying Resident and Immune Cell Injury Responses in the Human Kidney (1R21AG067335-01). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/9954680. Licensed CC0.

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