# Project III

> **NIH NIH P50** · MAGEE-WOMEN'S RES INST AND FOUNDATION · 2024 · $472,999

## Abstract

Abstract: Project 3: Somatic and germline gene therapy for male infertility
Infertility impacts 10-15% of couples in the United States and a male factor is implicated alone or in
combination with female factors in about 50% of cases. Infertility can be caused by hormonal, anatomical,
immunological or chromosomal deficiencies, diseases or medical treatments, but is frequently of unknown
origin (idiopathic). Idiopathic infertility is difficult to counsel and treatment options are limited. This program will
focus on patients with the most severe form of male infertility, nonobstructive azoospermia (NOA). Improved
knowledge about the genetic basis of NOA obtained in this program will aid in the counseling of infertile
couples; justify the development of diagnostic screens; and may lead to new patient-specific treatment options.
Project 3 will test the hypotheses that: 1) gene therapy/gene editing can be used to treat mouse models
of human NOA that exhibit somatic cell or germ cell dysfunction; 2) gene editing can be performed in
induced pluripotent stem cells (iPSCs) from mouse models of NOA, which can be differentiated into
transplantable germ cells or sperm; and 3) gene editing can be performed in NOA patient-derived
iPSCs that can be differentiated to primordial germ cell-like cells (PGCLCs). Project 3 will provide “gold
standard” reproductive phenotyping in mouse models with NOA-associated variants that are discovered in
Project 1 and validated with in vitro assays in Project 2. Mouse models that replicate the patient NOA
phenotype will be used for development of gene therapies for male infertility. For Sertoli cell defects, Aim 1 will
extend our previous results by demonstrating that adeno-associated viruses carrying corrective genes can be
introduced to Sertoli cells in vivo and restore spermatogenesis with low toxicity and without integrating in the
genome or modifying the germline. For germ cell defects, Aim 1 will prove the principle that NOA-associated
variants in spermatogonial stem cells (SSCs) can be corrected ex vivo with CRISPR/Cas9 or Prime editing and
transplanted to restore spermatogenesis and fertility in mouse models of NOA. Aim 2 will use CRISPR/Cas9 or
Prime editing to correct NOA-associated variants mouse iPSCs followed by differentiation to transplantable
PGCLCs or SSC-like cells (SSCLCs) to restore spermatogenesis and fertility. Aim 3 will use CRISPR/Cas9 or
Prime editing to correct validated NOA-associated variants in NOA patient-derived iPSCs followed by
differentiation to PGCLCs. This project will establish the safety and feasibility of gene therapies for male
infertility in mouse models and human cells to support future translation to the human clinic.

## Key facts

- **NIH application ID:** 10992993
- **Project number:** 2P50HD096723-06
- **Recipient organization:** MAGEE-WOMEN'S RES INST AND FOUNDATION
- **Principal Investigator:** Kyle Edwin Orwig
- **Activity code:** P50 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $472,999
- **Award type:** 2
- **Project period:** 2019-09-01 → 2028-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10992993

## Citation

> US National Institutes of Health, RePORTER application 10992993, Project III (2P50HD096723-06). Retrieved via AI Analytics 2026-05-27 from https://api.ai-analytics.org/grant/nih/10992993. Licensed CC0.

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