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Selective lysine‐specific demethylase 1 inhibitor, NCL1, could cause testicular toxicity via the regulation of apoptosis
- Source :
- Andrology
- Publication Year :
- 2020
- Publisher :
- Wiley, 2020.
-
Abstract
- Background Recent studies have shown that epigenetic alterations, such as those involving lysine‐specific demethylase 1 (LSD1), lead to oncogenic activation and highlight such alterations as therapeutic targets. However, studies evaluating the effect of LSD1 inhibitors on male fertility are lacking. Objectives We analyzed the potential toxicity of a new selective LSD1 inhibitor, N‐[(1S)‐3‐[3‐(trans‐2‐aminocyclopropyl)phenoxy]‐1‐(benzylcarbamoyl)propyl] benzamide (NCL1), in testes. Materials and methods Human testicular samples were immunohistochemically analyzed. Six‐week‐old male C57BL/6J mice were injected intraperitoneally with dimethyl sulfoxide vehicle (n = 15), or 1.0 (n = 15) or 3.0 (n = 15) mg/kg NCL1 biweekly. After five weeks, toxicity and gene expression were analyzed in testicular samples by ingenuity pathway analysis (IPA) using RNA sequence data and quantitative reverse transcriptase (qRT)–PCR; hormonal damage was analyzed in blood samples. NCL1 treated GC‐1, TM3, and TM4 cell lines were analyzed by cell viability, chromatin immunoprecipitation, flow cytometry, and Western blot assays. Results LSD1 was mainly expressed in human Sertoli and germ cells, with LSD1 levels significantly decreased in a progressive meiosis‐dependent manner; germ cells showed similar expression patterns in normal spermatogenesis and early/late maturation arrest. Histological examination revealed significantly increased levels of abnormal seminiferous tubules in 3.0 mg/kg NCL1–treated mice compared to control, with increased cellular detachment, sloughing, vacuolization, eosinophilic changes, and TUNEL‐positive cells. IPA and qRT–PCR revealed NCL1 treatment down‐regulated LSD1 activity. NCL1 also reduced total serum testosterone levels. Western blots of mouse testicular samples revealed NCL1 induced a marked elevation in cleaved caspases 3, 7, and 8, and connexin 43 proteins. NCL1 treatment significantly reduced GC‐1, but not TM3 and TM4, cell viability in a dose‐dependent manner. In flow cytometry analysis, NCL1 induced apoptosis in GC‐1 cells. Conclusions High‐dose NCL1 treatment targeting LSD1 caused dysfunctional spermatogenesis and induced caspase‐dependent apoptosis. This suggests the LSD1 inhibitor may cause testicular toxicity via the regulation of apoptosis.
- Subjects :
- Male
Urology
Endocrinology, Diabetes and Metabolism
Antineoplastic Agents
Apoptosis
Cell Line
Flow cytometry
Andrology
Mice
03 medical and health sciences
0302 clinical medicine
Endocrinology
Western blot
NCL1
Testis
medicine
Animals
Humans
Testosterone
Viability assay
Infertility, Male
Histone Demethylases
Sertoli Cells
toxicity in testis
030219 obstetrics & reproductive medicine
medicine.diagnostic_test
Chemistry
Prostatic Neoplasms
Original Articles
Spermatozoa
spermatogenesis
Mice, Inbred C57BL
Blot
Reproductive Medicine
Vacuolization
Hematologic Neoplasms
Toxicity
Original Article
lysine‐specific demethylase 1
Spermatogenesis
Subjects
Details
- ISSN :
- 20472927 and 20472919
- Volume :
- 8
- Database :
- OpenAIRE
- Journal :
- Andrology
- Accession number :
- edsair.doi.dedup.....026e9c641cdde526e58638300ee740be
- Full Text :
- https://doi.org/10.1111/andr.12846