Increased susceptibility to prostate cancer biomarkers in the offspring of male mouse progenitors with lifelong or early life exposure to high-fat diet

Abstract

Obesity exacerbates hormonal dysregulation, inflammation, and oxidative stress, factors associated with Prostate Cancer (PCa) development. The (epi)genetic influences of obesity may be transgenerationally transmitted, potentially impacting PCa susceptibility in the offspring of fathers with obesity. Thus, we studied the impact of early-life or lifelong exposure to a high-fat diet (HFD) on PCa biomarkers [Homeobox B13 (HOXB13) and the Androgen Receptor (AR)] and their correlation with obesity-related markers. Furthermore, we focused on the offspring's PCa biomarkers outcomes and explored their potential link with paternal diet. A transgenerational Mus musculus model was established, with F0 males exposed to different diets (200 days): standard chow, lifelong HFD (HFD), and transient diet (60 days HFD, plus 140 days of standard chow) (HFD_t). AR expression in the prostates was unaffected, whereas HOXB13, Fat Mass and Obesity Associated gene (FTO), and Tumor Necrosis Factor-Alpha (TNF-α) expression decreased in the F1 HFD_t group. HOXB13 and AR prostate expression were positively correlated. There was also a positive correlation between FTO prostate expression and PCa biomarkers, and between TNF-α expression and FTO and PCa biomarkers. HOXB13 promoter methylation levels were unaffected, however, were positively correlated with FTO and HOXB13 expression. Finally, protein nitration remained unchanged in the prostates, while lipid peroxidation was increased in the F0 HFD group and decreased in the F1 and F2 HFD and HFD_t groups. Our study highlights the intergenerational interplay between obesity-related factors and PCa biomarkers, suggesting that offspring of male progenitors subjected to HFD may face an increased risk for elevated PCa biomarkers expression.

Keywords: Epigenetic; Obesity; Prostate cancer; Transgenerational.

Fig. 1

Schematic representation of the transgenerational animal model

Fig. 2

Identification of PCa biomarkers mRNA in mouse prostate tissue subjected to standard chow by conventional PCR. Conventional PCR was used to identify the presence of Androgen Receptor (A) and Homeobox B13 (B) transcripts in mouse prostate tissue. A cDNA-free sample was used as a negative control. A mouse liver tissue sample was used as a positive control

Fig. 3

Expression of AR (a, b, c), HOXB13 (d, e, f), FTO (g, h, i), and TNF-α (j, k, l) in the prostate of mice fed with standard chow (CTRL) or lifelong high-fat diet (HFD) or only early life HFD (HFD_t) in F0 and on their offspring (F1 and F2). Data shows the expression of AR, HOXB13, FTO, and TNF-α in the prostate of mice (F0, F1, and F2). The expression was determined by qPCR. Results are expressed as mean ± SD, (n = 4–8 for each bar). Significantly different results (p < 0.05) are as indicated: * relative to HFD

Fig. 4

Identification of methylation-specific primers, HOXB13 unmethylated and HOXB13 methylated, in mice prostate tissue by MSP (a), methylation levels of HOXB13 promoter (b) and correlation between HOXB13 promoter methylation levels and FTO (c) and HOXB13 (d) expression in the prostate of mice fed with standard chow (CTRL) or lifelong high-fat diet (HFD) or early life HFD (HFD_t) in F0 and on their offspring (F1, and F2). Panel a shows the identification of the presence of unmethylated HOXB13 and methylated HOXB13 transcripts in mice prostate tissue through MSP. A cDNA-free sample was used as the negative control. Panel b shows the methylation levels of the HOXB13 promoter in the prostate of mice. The expression was determined by qMSP. Results are expressed as mean ± SD (n = 5 for each graph bar), except for the group HFD_t in F2 (n = 4). Panel c shows the correlation between HOXB13 promoter methylation levels and FTO expression in the prostate of F0 (n = 13) and F1 (n = 14) mice. Panel d shows the correlation between HOXB13 promoter methylation levels and HOXB13 expression in the prostate of F0 (n = 13) mice. Outliers were identified by ROUT (Q = 10%). All P values < 0.05 were considered statistically significant

Fig. 5

Evaluation of protein nitration (a) and lipid peroxidation (b) in the prostate of mice fed with standard chow (CTRL) or lifelong high-fat diet (HFD) or early life HFD (HFD_t) of progenitors in F0 and on their offspring (F1, and F2). Data shows the analysis of protein nitration (a) and lipid peroxidation (b) in the prostate of mice of F0, F1, and F2. The expression was determined by Slot-Blot. Results are expressed as mean ± SD (n = 6 for each graph bar), except for the groups CTRL (n = 4) and HFD_t (n = 3) in F2. Significantly different results (p < 0.05) are as indicated: a relative to CTRL, b relative to HFD

Fig. 6

Venn diagram illustrating the proteins present in mouse (a) and human (b) prostate that interact with FTO and/or HOXB13 and protein-protein interactions network of the identified FTO and HOXB13 interactors in human prostate (c). The node size is proportional to the number of interactions