SummaryA link between elevated luteinizing hormone (LH) levels, GATA-4 and LH receptor (LHCGR) expression and gonadotropin-dependent adrenocortical tumorigenesis in humans and mice has been shown. To assess the mechanistic tumorigenic interrelationships between these factors, we transgenically expressed Gata4 under the 21-hydroxylase promoter (Cyp21a1, 21-OH) in C57Bl/6N mice. There was a gradual age-dependent increase of GATA-4 expression only in 21-OH-GATA-4 (TG) female adrenals, in association with slowly progressing neoplasia of non-steroidogenic spindle-shaped A cells in the subcapsular cortex. Gonadectomy (GDX), apparently through direct action of elevated serum LH, markedly enhanced the adrenocortical neoplasia, which now also appeared in GDX TG males. The neoplastic areas of the post-GDX TG adrenals contained, besides A cells, larger lipid-laden, steroidogenically active and LHCGRpositive B cells. Prolonged (.10 months) exposure to elevated post-GDX LH levels resulted in formation of adrenocortical adenomas in the TG mice. Intact and GDX TG mouse adrenals displayed elevated FOG-2 and decreased GATA-6 expression. Additionally, increased expression/activation of components of the Inhbb-Acvr2a-Acvr1c-Smad2/3 signaling system was observed in 12-month-old GDX TG adrenals. Our findings show that two distinct GATA-4-dependent populations of neoplastic adrenocortical cells form: non-steroidogenic LH-independent A cells and steroidogenic LH-dependent B cells.

Hypothyroidism has been shown to be associated with a reduction in serum testosterone level in males. This reduction in testosterone is reversible by thyroxine replacement therapy. However, to the best of our knowledge, it is not yet known, whether a similar reduction in serum testosterone level is observed in subclinically hypothyroid males [thyroid-stimulating hormone (TSH) < 10 mIU/L] in whom the benefits of thyroxine replacement therapy are still controversial. Our goal was to investigate the putative connections between subclinical hypothyroidism and the circulating levels of gonadotrophins and gonadal steroids in males (mean age +/- SEM, 34.67 +/- 1.52 years; ranging from 20 to 54 years). The serum samples from patients showing normal euthyroid and subclinical hypothyroid profiles (TSH < 10 mIU/L) were further analysed for the levels of luteinizing hormone, follicle-stimulating hormone, prolactin, testosterone, sex hormone-binding globulin, progesterone and oestradiol. Subclinical hypothyroidism was associated with a decrease in the levels of serum testosterone and its precursor progesterone. The data suggest that serum testosterone declines because of the non-availability of its precursor progesterone. The level of oestradiol was similar in both the groups, suggesting a greater conversion rate of testosterone to oestradiol in subclinically hypothyroid males, in order to maintain the oestradiol levels. Prolactin levels were slightly but significantly increased in subclinical hypothyroidism. To the best of our information this is a novel report, which shows a direct association between subclinical hypothyroidism and hypoandrogenaemia. Testosterone deficiency and its symptoms should be kept in view while managing subclinical hypothyroidism in male patients. Further studies are needed in order to reveal the physiological and molecular mechanisms leading to hypoandrogenaemia in subclinical hypothyroidism (TSH < 10 mIU/L).

Inconsistent alterations in gonadal steroidogenesis and pituitary functions have been reported in hypothyroid males. We have compared the lipid and endocrine profiles of the euthyroid and hypothyroid [thyroid-stimulating hormone (TSH) >100 mIU l(-1)] males. Hypothyroidism was found to be associated with an increase in the circulating level of total cholesterol and LDL-cholesterol (LDL-C) and a reduction in the levels of progesterone and testosterone, without any change in the serum levels of oestradiol and gonadotrophins. The failure of gonadotrophins to rise could be accounted by a normal level of serum oestradiol in the hypothyroid male. A mild hyperprolactinaemia was also noted in the hypothyroid patients. The reduction in serum testosterone level could be explained by (i) a reduced uptake of LDL-C by the Leydig cells and thereby a reduction in the synthesis of progesterone and consequentially testosterone, (ii) a further reduction in the rate of conversion of progesterone to testosterone, (iii) a higher rate of conversion of testosterone to oestradiol, (iv) a decrease in serum triiodothyronine and (v) hyperprolactinaemia. Rise in TSH needs to be investigated as a cause of the suppression of gonadal steroidogenesis.

Lytic peptide Hecate (23-amino acid (AA)) fused with a 15-AA fragment of human chorionic gonadotropin-b (CG-b), Hecate-CGb conjugate (H-CGb-c) selectively binds to and destroys tumor cells expressing LH/chorionic gonadotropin receptor (Lhcgr). Transgenic mice (6.5 month old) expressing SV40 T-antigen under the inhibin-a promoter (inha/Tag) presenting with Lhcgr expressing adrenal tumors were treated either with H-CGb-c, GnRH antagonist (GnRH-a), estradiol (E 2 ; only females) or their combinations for 1 month. We expected that GnRH-a or E 2 in combination with H-CGb-c could improve the treatment efficacy especially in females by decreasing circulating LH and eliminating the potential competition of serum LH with the H-CGb-c. GnRH-a and H-CGb-c treatments were successful in males (adrenal weights 14G2.8 mg and 60G26 vs 237G59 mg in controls; P!0.05). Histopathologically, GnRH-a apparently destroyed the adrenal parenchyma leaving only the fibrotic capsule with few necrotic foci. In females, H-CGb-c was totally ineffective, whereas GnRH-a (19G5 mg) or E 2 (77G50 mg) significantly reduced the adrenal weights compared with controls (330G70 mg). Adrenal morphometry, cell proliferation markers, post-treatment suppression of serum progesterone, and quantitative RT-PCR of GATA-4, Lhcgr, and GATA-6 further supported the positive outcome. H-CGb-c selectively killed the Lhcgr expressing tumor cells, whereas GnRH-a blocked tumor progression through gonadotropin suppression, emphasizing the gonadotropin dependency of these adrenocortical tumors. If extrapolated to humans, H-CGb-c could be considered for the treatment of gonadotropin-dependent adrenal tumors in males, whereas in females gonadotropin suppression, but not H-CGb-c, would work better.

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