Protein kinase A drives paracrine crisis and WNT4-dependent testis tumor in Carney complex
Cyril Djari, … , Antoine Martinez, Anne-Marie Lefrançois-Martinez
Cyril Djari, … , Antoine Martinez, Anne-Marie Lefrançois-Martinez
Published December 1, 2021
Citation Information: J Clin Invest. 2021;131(23):e146910. https://doi.org/10.1172/JCI146910.
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Research Article Endocrinology Reproductive biology

Protein kinase A drives paracrine crisis and WNT4-dependent testis tumor in Carney complex

Abstract

Large-cell calcifying Sertoli cell tumors (LCCSCTs) are among the most frequent lesions occurring in male Carney complex (CNC) patients. Although they constitute a key diagnostic criterion for this rare multiple neoplasia syndrome resulting from inactivating mutations of the tumor suppressor PRKAR1A, leading to unrepressed PKA activity, LCCSCT pathogenesis and origin remain elusive. Mouse models targeting Prkar1a inactivation in all somatic populations or separately in each cell type were generated to decipher the molecular and paracrine networks involved in the induction of CNC testis lesions. We demonstrate that the Prkar1a mutation was required in both stromal and Sertoli cells for the occurrence of LCCSCTs. Integrative analyses comparing transcriptomic, immunohistological data and phenotype of mutant mouse combinations led to the understanding of human LCCSCT pathogenesis and demonstrated PKA-induced paracrine molecular circuits in which the aberrant WNT4 signal production is a limiting step in shaping intratubular lesions and tumor expansion both in a mouse model and in human CNC testes.

Authors

Cyril Djari, Isabelle Sahut-Barnola, Amandine Septier, Ingrid Plotton, Nathanaëlle Montanier, Damien Dufour, Adrien Levasseur, James Wilmouth Jr., Jean-Christophe Pointud, Fabio R. Faucz, Crystal Kamilaris, Antoine-Guy Lopez, Florian Guillou, Amanda Swain, Seppo J. Vainio, Igor Tauveron, Pierre Val, Hervé Lefebvre, Constantine A. Stratakis, Antoine Martinez, Anne-Marie Lefrançois-Martinez

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Figure 8

PKA activation induces mTOR, MAPK, and hypoxia pathways in tumor of human LCCSCTs.

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PKA activation induces mTOR, MAPK, and hypoxia pathways in tumor of huma...
(A) GO enrichment scores of tumor development–associated pathways based on gene list specifically deregulated in 2-month-old prKO testis compared with srKO and lrKO testes. (B and C) GSEA of microarray gene expression data showing positive enrichment of hallmark-MTORC1 signaling gene list (B) and KEGG-MAPK signaling pathway gene list (C) in 2-month-old prKO testis compared with WT. (D) Heatmap representing the median-centered expression of significantly deregulated growth factors and associated receptors (adj. P < 0.05) in 2-month-old prKO (n = 3–4) compared with WT (n = 4). (E) RT-qPCR analysis of Egf, Fgf2, Igf1, Tgfb1, Tgfb2, and Tgfb3 transcripts in 4-month-old WT, prKO, and prwKO testes. One-way ANOVA was followed by Tukey’s multiple-correction test, Welch’s 1-way ANOVA was followed by Games-Howell multiple-correction test, and Kruskal-Wallis test was followed by Dunn’s multiple-correction test. (F) Western blot analysis of p4EBP1, pS6K, and pERK accumulation in 4-month-old WT, prKO, and prwKO testes. (G) Phosphorylated protein quantification over total proteins. Statistical analysis was performed using Student’s t test. (H) Immunohistochemical detection of p4EBP1 in 4-month-old WT and prKO testes. (I and J) Immunohistochemical detection of p4EBP1 and pERK in CNC-LCCSCTs. Insets at the top right (I) correspond to areas adjacent to the tumor characterized by low p4EBP1 staining. (K) GSEA of microarray gene expression data showing positive enrichment of hallmark-hypoxia gene list (genes upregulated in response to hypoxia) in 2-month-old prKO testis compared with WT. (L) Hif1a, Hif1b, and Hif2a transcript analysis in 4-month-old WT and prKO testes. Statistical analysis was performed using Welch’s or Student’s t test. (M) Immunohistochemical detection of HIF2A in CNC-LCCSCTs. Bars represent the mean per group ± SD. Scale bars: 100 μm. Original magnification, ×1.60 (left insets, I, J, and M); ×0.80 (right insets, I and M). *P < 0.05; ***P < 0.001.