Ghosh-Choudhary, Shohini
(2025)
Rational Identification of Novel Senolytic Therapies Revealed by a Genome-Wide CRISPR Screen.
Doctoral Dissertation, University of Pittsburgh.
(Unpublished)
Abstract
By the year 2050, the largest demographic in the world will be individuals over 60 years of age2-4. Accompanying this demographic shift, there is an increased focus on aging and healthy longevity. With aging known as a risk factor in a variety of diseases, the biology of aging has become pertinent to the scientific understanding of these diseases. The goal of many researchers in this area is to be able to treat the underlying pathological mechanisms. One of the fundamental processes contributing to pathological aging is senescence, the irreversible growth arrest of cells in reaction to a stressor. The hallmarks of aging have listed senescence as one keys to understand aging biology since 20136,7. Senescent cells are being linked to a variety of diseases from atherosclerosis to osteoarthritis9-14. While the process of senescence safeguards against the transformation of damaged cells into cancerous cells, the persistence of these senescent cells has proved to be deleterious15,16. These senescent cells are notoriously difficult to kill once they persist in tissues unless they are cleared by a competent immune system9,13,14. These cells contribute to inflammatory paracrine signaling through secreted factors9,17.
With our understanding of senescence deepening, many have sought to develop drug candidates that can kill these persistent senescent cells. Some of the first senolytic drugs developed are now being tested in clinical trials to determine their efficacy in a variety of diseases9. These first generations of drugs sought to tip the balance of anti-apoptotic mechanisms in senescent cells towards a more pro-apoptotic mechanism9,17,19. These drugs have proven to have a narrow therapeutic index making their efficacy in the clinic difficult to assess without unwanted side effects30,60. To develop better senolytics, or drugs to target senescent cells, there is a need to develop rational approaches to better understanding senescent cell biology and to exploit any underlying vulnerabilities. In this study, we perform a novel whole genome CRISPR screen to select for targets that lead to the death of senescent cells without affecting the growth of healthy replicating cells. From this screen, we go on to validate PARP16 as a selectively lethal target in a variety of senescent cell types and following a variety of senescent cell triggers. Finally, we show that two small molecule inhibitors of PARP16 are able to specifically kill senescent cells. Using this platform, we show that systematic unbiased assessment of all genes in the genome allows the rational design of selective senolytic drugs.
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Details
| Item Type: |
University of Pittsburgh ETD
|
| Status: |
Unpublished |
| Creators/Authors: |
| Creators | Email | Pitt Username | ORCID  |
|---|
| Ghosh-Choudhary, Shohini | skg46@pitt.edu | skg46 | |
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| ETD Committee: |
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| Date: |
24 February 2025 |
| Date Type: |
Publication |
| Defense Date: |
10 September 2024 |
| Approval Date: |
24 February 2025 |
| Submission Date: |
17 November 2024 |
| Access Restriction: |
2 year -- Restrict access to University of Pittsburgh for a period of 2 years. |
| Number of Pages: |
128 |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
School of Medicine > Cellular and Molecular Pathology |
| Degree: |
PhD - Doctor of Philosophy |
| Thesis Type: |
Doctoral Dissertation |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
Senescence
Senolytic
PARP16
Aging
CRISPR Screen |
| Date Deposited: |
24 Feb 2025 17:17 |
| Last Modified: |
24 Feb 2025 17:17 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/47087 |
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