# 3D Culture Systems Of Urine-Derived Stem Cell For NTRI-Induced Mitotoxicity Assessment

> **NIH NIH R21** · WAKE FOREST UNIVERSITY HEALTH SCIENCES · 2020 · $193,750

## Abstract

Summary
Antiretroviral therapy (ART) has significantly reduced HIV-related morbidity and mortality. However, therapeutic
life expectancy for individuals with HIV
increased, the long-term safety of ART has gained increasing attention. Thus, it is highly important to
the long-term safety of antiretroviral regimens. Human hepatocyte cell lines in 2D culture are most
used to evaluate short-term mitochondrial toxicity (MtT) induced by ART. However, these
cannot be used in detection of ART-induced chronic MtT because they cannot survive more than
days during toxicity testing. In addition, antiviral drug toxicity screening often requires expensive primary
such as peripheral blood monocytes in 2D culture systems. More than 90% of drugs that pass through in
2D culture preclinical studies fail to meet the desired efficacy or safety margins required in subsequent
trials. Clearly, 2D cultures have contributed to the poor predictive power of MtT screening assays in
. Although long-term toxicity tests are often performed in animal models, the high rates of MtT observed in
earlier clinical trials suggest that animal toxicology studies may not be suitable for predicting MtT of antiviral
intended for human use. Apparently, there are no adequate 2D cell culture or animal models for late MtT
for preclinical drug development. Hence, there is an urgent need to develop more toxicologically
and clinically predictive in vitro assays to assess compounds for the potential of late MtT. We were the
to demonstrate that stem/progenitor cells exist in human urine, i.e., urine-derived stem cells (USC). These
can be obtained using simple, non-invasive and low-cost procedures. USC express telomerase activity
possess robust proliferative potential. We recently revealed that 3D culture of USC provides a long-term,
microenvironmen for cell growth and proliferation, mimicking in vivo conditions, which may have
to improve the predictive outcome of preclinical antiviral drug studies. Thus,
is to develop 3D culture systems of human USC for ART-induced MtT testing. We hypothesize that human
USC maintain telomerase activity and mitochondrial function in 3D organoids, which considerably extends the
life of cell culture systems, enabling long-term assessment of late ART-induced MtT . To test this hypothesis,
we propose the following: Aim 1. Assess the stemness properties and mitochondrial function of human USC in
3D organoids over long-term culture, compared to 2D culture of USC; Aim 2. Determine the cytotoxicity,
inhibition of Pol-γ, mitochondrial DNA content and MtT profiles of 9 antiretroviral drugs in 3D USC cultures. We
that the antiretroviral drugs tested will exhibit effects on MtT in the 3D culture systems and be ranked
to their effective concentrations based on in vitro MtT assay. Therefore, the use of patients-derived
cells from urine to generate 3D culture assay offers a promising tool for antiretroviral drug development
personalized medicine in the assessment of MtT of anti-HIV ...

## Key facts

- **NIH application ID:** 10083026
- **Project number:** 1R21AI152832-01A1
- **Recipient organization:** WAKE FOREST UNIVERSITY HEALTH SCIENCES
- **Principal Investigator:** YUANYUAN no ZHANG
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $193,750
- **Award type:** 1
- **Project period:** 2020-07-10 → 2022-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10083026

## Citation

> US National Institutes of Health, RePORTER application 10083026, 3D Culture Systems Of Urine-Derived Stem Cell For NTRI-Induced Mitotoxicity Assessment (1R21AI152832-01A1). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10083026. Licensed CC0.

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