# Methods for detection of dynamic intracellular signals in single adult spermatogonial stem cells

> **NIH NIH R21** · WEILL MEDICAL COLL OF CORNELL UNIV · 2024 · $210,086

## Abstract

Abstract
Homeostasis and differentiation of adult mammalian spermatogonial stem cells (SSCs) depend on tightly
regulated signal transduction through canonical pathways (e.g., RAS/ERK MAPK and PI3K/AKT). Disruption of
these pathways by pathogenic de novo mutations leads to changes in cell fitness and paternal age-related
accumulation of mutant SSC clones that generate craniofacial disorders in children. Recent studies in other
cell types show that dynamic patterns of signaling (on the scale of minutes to hours) are central to phenotypic
behavior of cells. Yet, the data for signaling requirements in SSCs until now have been derived from binary,
static measures, representing a major barrier to reconciling paradoxical observations regarding the roles of
specific pathways in SSCs. Furthermore, it has been difficult or impossible to link upstream signals (i.e., growth
factors) with specific downstream effectors due to parallel pathway activation, both in the context of normal
SSCs and those with pathogenic mutations. The general goal of this proposal is to develop novel methodology
to reveal how information is encoded by dynamic signaling patterns in SSCs, using kinase translocation
reporter technology (KTR), which enables quantitative, real-time measurement of pathway activity. As proof-
of-principle, we will apply this strategy to probe ERK MAPK and PI3K/AKT signaling driven by canonical
receptor tyrosine kinases that are closely linked to self-renewal of SSCs, both in wild type cells and those with
mutations in genes that drive paternal age-associated craniofacial disorders. However, we anticipate our
results will be useful across a variety of pathways for revealing how extrinsic signals from the extracellular
environment are transmitted internally, dynamically encoded, and modified by network cross talk.

## Key facts

- **NIH application ID:** 10817956
- **Project number:** 5R21HD107212-02
- **Recipient organization:** WEILL MEDICAL COLL OF CORNELL UNIV
- **Principal Investigator:** Todd R Evans
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $210,086
- **Award type:** 5
- **Project period:** 2023-04-01 → 2025-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10817956

## Citation

> US National Institutes of Health, RePORTER application 10817956, Methods for detection of dynamic intracellular signals in single adult spermatogonial stem cells (5R21HD107212-02). Retrieved via AI Analytics 2026-05-27 from https://api.ai-analytics.org/grant/nih/10817956. Licensed CC0.

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