OR04-01 Single Nucleus RNA Sequencing Reveals Diet Induced Obesity And SGLT2 Inhibitor Therapy Modulate the Testes Transcriptome In Male Mice

Disclosure: B. Ozturk: None. V. Efthymiou: None. W. Ali: None. R. Ferraz-Bannitz: None. P. Erika: None. V. Navarrete: None. J.M. Dreyfuss: None. H. Pan: None. M. Patti: Consulting Fee; Self; AstraZeneca, MBX-Biosciences, Hanmi Pharmaceutical. Other; Self; DSMB: Fractyl Health, Inc. Paternal environmental exposures during postnatal life can influence health of their offsprings, modulating risk of metabolic disease. We have previously shown that pharmacologic interventions to improve the health of fathers may also reduce metabolic disease risk of offsprings. To identify potential mechanisms responsible for paternally-mediated intergenerational metabolic disease risk, we analyzed the transcriptome in testes using single-nucleus RNA sequencing. C57BL/6J male mice, age 8 weeks, were treated with low fat (10% fat) or high fat diet (HFD, 60% fat) diet to induce obesity and hyperglycemia; after 4 weeks, HFD-fed mice were randomized to continued HFD or HFD containing the SGLT2 inhibitor canagliflozin (250 mg/ kg/ day) (CANA). After 20 weeks, we dissected testes for single nucleus RNA sequencing (LFD, HFD, CANA n=3,4,4 respectively). Unsupervised clustering revealed 35 distinct clusters. We employed supervised cell-calling and identified 13 distinct canonical testis-residing cell types and 2 novel subtypes. The magnitude of differential expression between groups was most pronounced in Leydig cells, pachytene-stage cells, and round spermatids, suggesting that the transcriptome of particular cell types in testis is more robustly affected in response to obesity. Ontology analysis of differentially expressed genes (DEG) in those 3 cell types (FDR≤0.1 for input genes and p≤0.05 for enrichment analysis) revealed that downregulated genes in HFD as compared to LFD were enriched in cytoskeletal element organization, posttransciptional and posttranslational modifications respectively. Importantly, upregulated genes in CANA compared to HFD were also enriched in similar ontology terms. In addition, ontology analysis of DEGs in CANA compared to HFD showed enrichment in mitochondrial oxidative phosphorylation in Leydig cells and round spermatids which suggests unique transcriptomic changes driven by canagliflozin in these cell types. Thus, diet-induced obesity has striking effects to remodel the testicular transcriptome, with substantial reversal by SGLT2i. Transcriptomic differences induced by high fat diet and reversed by SGLT2i may reveal molecular mechanisms that mediate the reduction in risk of metabolic disease to offspring. Presentation: Thursday, June 15, 2023