Quantification of the Leydig cell compartment in testicular biopsies and association with biochemical Leydig cell dysfunction in testicular cancer survivors

Abstract

A simple histological method to evaluate the Leydig cell compartment is lacking. We aimed to establish such a method and to investigate if Leydig cell hyperplasia of the biopsy contralateral to the tumour-bearing testicle in patients with testicular germ cell cancer is associated with biochemical signs of Leydig cell dysfunction after long-term follow-up. A case group of 50 long-term testicular germ cell cancer survivors without human chorionic gonadotropin elevation, 10 testicular germ cell cancer patients with elevated human chorionic gonadotropin and 10 controls without testicular malignancy were included. For each subject, 2-4 representative sections from their testicular biopsies were selected for analysis. Using the image processing program ImageJ (V.1.48, NIH), an area with a minimum of 50 tubules was selected and delineated (total selected area) and the total Leydig cell area was calculated by adding up every delineated Leydig cell group within the total selected area. Four different methods were tested for the ability to quantify the Leydig cell compartment. In the 50 testicular germ cell cancer survivors, associations between the area of the Leydig cell compartment and serum levels of testosterone and luteinising hormone were investigated using linear regression analysis. The Leydig cell compartment was best quantified by the total Leydig cell area/total selected area index, which was significantly larger in the human chorionic gonadotropin-positive patients than in controls (P = 0.00001). In the 50 human chorionic gonadotropin-negative testicular germ cell cancer survivors, increasing total Leydig cell area/total selected area was significantly associated with decreased levels of total testosterone and decreased total testosterone/luteinising hormone ratio after a median of 9-year follow-up. In conclusion, a new simple method, total Leydig cell area/total selected area, was established to estimate the Leydig cell compartment in testicular biopsies. The index identified Leydig cell hyperplasia in the contralateral biopsy in patients with testicular germ cell cancer, and it was associated with long-term biochemical Leydig cell dysfunction. Although in testicular germ cell cancer survivors, the clinical value is limited because the contralateral biopsies are not commonly available, we propose a closer andrological follow-up in any patient with an increased total Leydig cell area/total selected area index.

Clinical presentation, management and follow-up of 83 patients with Leydig cell tumors of the testis: a prospective case-cohort study

“…It is hypothesized that many cases of abnormal spermatogenesis, cryptorchidism, penile malformations, and testicular cancer may have a common etiology, such as an irreversible developmental disorder originating in early fetal life and resulting in TDS (Joensen et al , 2008). Leydig cell hyperplasia is a frequent finding in patients with impaired spermatogenesis and other TDS-related disorders, often in the form of large clusters called micronodules (Joensen et al , 2008; Soerensen et al , 2016; Tarsitano et al , 2018). Leydig cell micronodules, defined as more than 15 Leydig cells in a cross-section, have previously been associated with a low testosterone/LH ratio, reflecting an endocrine dysfunction (Joensen et al , 2008).…”

Transcriptome analysis of human Leydig cell tumours reveals potential mechanisms underlying its development

“…Of note, Leydig cells can control the steroidogenesis and VEGF production under the homeostatic scope of the testicular temperature (Goyal & Saini, 2018). Leydig cell hyperplasia is a frequent finding in patients with impaired spermatogenesis and another testicular dysgenesis syndrome (TDS)‐related disorder (Tarsitano et al, 2018) and has previously been associated with a low testosterone/lutropin ratio. Besides lutropin that in pathological conditions promotes Leydig cell proliferation, human endocrine gland derived vascular endothelial growth factor (EG‐VEGF) secretion can induce the growth of Leydig cell tumours (Samson et al., 2004).…”

“…Of note, Leydig cells can control the steroidogenesis and VEGF production under the homeostatic scope of the testicular temperature (Goyal & Saini, 2018). Leydig cell hyperplasia is a frequent finding in patients with impaired spermatogenesis and another testicular dysgenesis syndrome (TDS)-related disorder (Tarsitano et al, 2018) and has previously been associated with a low testosterone/lutropin ratio.…”

Serum Concentrations and Gonadal Expression of INSL3 in Eighteen Males With 45,X/46,XY Mosaicism

“…Leydig cell hyperplasia was quantified by determining the “Total Leydig Cell Area (TLCA)/Total Selected Area (TSA)” as described by Tarsitano et al. with a few modifications ( 27 ). In brief, quantification was conducted on the sections stained for CYP11A1 expression and the entire area of the testicular biopsy was included, i.e.…”