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Studies in the past decade have shown that lipid droplets stored in liver cells under starvation are encapsulated by autophagosomes and fused to lysosomes via the endocytic system. Autophagy responds to a variety of environmental factors inside and outside the cell, so it has a complex signal regulation network. To this end, we first explored the role of Hedgehog (Hh) in autophagy and lipid metabolism. Treatment of normal mouse liver cells with SAG and GDC-0449 revealed elevated phosphorylation of AMP-activated protein kinase (AMPK) and increased lipidation of LC3. SAG, and GDC-0449 were agonist and antagonist of Smoothened (Smo) in canonical Hh pathway, respectively, but they played a consistent role in the regulation of autophagy in hepatocytes. Moreover, SAG and GDC-0449 did not affect the expression of glioma-associated oncogene (Gli1) and patched 1, suggesting the absence of canonical Hh signaling in hepatocytes. We further knocked down the Smo and found that the efects of SAG and GDC-0449 disappeared, indicating that the non-canonical Smo pathway was involved in the regulation of autophagy in hepatocytes. In addition, SAG and GDC-0449 promoted lipid degradation and inhibited lipid production signals. Knockdown of Smo slowed down the rate of lipid degradation rather than Sufu or Gli1, indicating that Hh signaling regulated the lipid metabolism via Smo. In summary, activates AMPK via Smo to promote autophagy and lipid degradation. Key words: Hh signaling, Smo, AMPK, autophagy, lipid degradation, Introduction Hedgehog (Hh) signaling, which was first identified in Drosophila, plays a vital role in embryonic development and adult stem cell self-renewal (Nusslein-Volhard and Wieschaus 1980; Ingham and McMahon 2001; [...]

Metformin, an antidiabetic drug, has recently been repositioned in the treatment of several nondiabetic disorders, including reproductive disorders such as polycystic ovarian syndrome, where it improves endometrial functions. In vitro studies employing supratherapeutic concentrations (5-20 mmol/L) of metformin have reported antiproliferative effects on endometrial epithelial and stromal cells. However, animal and human studies have revealed that therapeutic serum concentrations of metformin range between 20 and 70 [micro]mol/L. In the present study, the effect of therapeutic concentrations of metformin was studied on endometrial epithelial cells (EECs). Therapeutic concentrations of metformin induced proliferation in Ishikawa and HEC-1A cells. The proliferation of EECs was found to be mammalian target of rapamycin (mTOR) dependent. Interestingly, therapeutic metformin concentrations were not able to activate the classical AMP-activated protein kinase (AMPK) signaling. On the contrary, supratherapeutic metformin concentration (10 mmol/L) inhibited mTOR and activated AMPK signaling. Microarray analysis of metformin-treated HEC-1A cells revealed dose-dependent differential effects on biological pathways associated with translation, ribosomal RNA processing, mitochondrial translation, and cell proliferation. Therapeutic concentrations of metformin upregulated mitochondrial number as demonstrated by increased MitoTracker[TM] Red staining and enhanced succinate dehydrogenase expression; however, higher concentration (10 mmol/L) abrogated the same. Our results suggest that therapeutic concentrations of metformin augment mitochondrial strength and induce mTOR-dependent endometrial cell proliferation. Key words: metformin, endometrium, proliferation, microarray, mitochondria, Introduction Metformin, primarily an antidiabetic drug, has been repositioned for various other nondiabetic purposes in recent years. Metformin, which was discovered initially to treat type 2 diabetes, has been shown [...]

The ability to change an organism's DNA through gene editing is of great importance for the prevention and treatment of genetic and acquired diseases. Rapid progress has been made during the last decade due to the discovery and refinement of CRISPR/Cas9 as an accurate, fast, and reliable genome editing technique. In this issue of the JCI, Lebek et al. present the culmination from a line of work in the Olson laboratory focused on in vivo gene editing of CAMK2D. The paper presents a combined state-of-the-art gene therapy approach that demonstrates how gene therapy can yield cardioprotection in a mouse model and takes notable steps toward potential applicability in patients., CAMK2D The CAMK2D gene encodes calcium/ calmodulin dependent protein kinase II delta (CaMKII[delta]), one of the major CaMKII isoforms and the predominant isoform in the heart. CaMKII[delta] contributes to the [...]

Cardiovascular diseases are the most common cause of worldwide morbidity and mortality, highlighting the necessity for advanced therapeutic strategies. [Ca.sup.2+]/calmodulin-dependent protein kinase IIS (CaMKII[beta]) is a prominent inducer of various cardiac disorders, which is mediated by 2 oxidation-sensitive methionine residues within the regulatory domain. We have previously shown that ablation of CaMKII[beta] oxidation by CRISPR-Cas9 base editing enables the heart to recover function from otherwise severe damage following ischemia/reperfusion (IR) injury. Here, we extended this therapeutic concept toward potential clinical translation. We generated a humanized CAMK2D knockin mouse model in which the genomic sequence encoding the entire regulatory domain was replaced with the human sequence. This enabled comparison and optimization of two different editing strategies for the human genome in mice. To edit CAMK2D in vivo, we packaged the optimized editing components into an engineered myotropic adeno-associated virus (MyoAAV 2A), which enabled efficient delivery at a very low AAV dose into the humanized mice at the time of IR injury. CAMK2D-edited mice recovered cardiac function, showed improved exercise performance, and were protected from myocardial fibrosis, which was otherwise observed in injured control mice after IR. Our findings identify a potentially effective strategy for cardioprotection in response to oxidative damage., Introduction Cardiovascular diseases, in particular coronary artery heart disease with subsequent myocardial infarction, are the most common cause of death worldwide, highlighting the necessity for advanced therapeutic strategies (1). Chronic [...]

The A-kinase anchoring protein 5 (AKAP5) has a variety of biological activities. This study explored whether AKAP5 was involved in cardiomyocyte apoptosis induced by hypoxia and reoxygenation (H/R) and its possible mechanism. H9C2 cells were used to construct an H/R model in vitro, followed by AKAP5 overexpression. Flow cytometry was performed to determine the rate of cardiomyocyte apoptosis. Phosphorylation of phospholamban (PLN), sarcoplasmic/endoplasmic reticulum calcium ATPase 2a (SERCA2a), and apoptosis-related proteins was determined by western blotting. Immunofluorescence staining and immunoprecipitation were performed to detect the distribution and interaction between AKAP5, protein kinase A (PKA), and PLN. After H/R induction, H9C2 cells exhibited significantly reduced AKAP5 protein expression. Upregulation of AKAP5 promotes cell survival and significantly reduces lactate dehydrogenase (LDH) levels and apoptosis rates in H9C2 cells. In addition, the overexpression of AKAP5 was accompanied by the activation of the PLN/SERCA2a signaling pathway and a reduction in apoptosis. Immunofluorescence staining and immunoprecipitation revealed that AKAP5 colocalized and interacted with PLN and PKA. Interestingly, St-Ht31, an inhibitory peptide that disrupts AKAP interactions with regulatory subunits, inhibits the effect of AKAP5 overexpression on H/R-induced apoptosis in H9C2 cardiomyocytes. AKAP5 overexpression alleviated H/R-induced cardiomyocyte apoptosis possibly by anchoring PKA to mediate the PLN/SERCA pathway, suggesting that AKAP5 is a potential therapeutic target for the prevention and treatment of ischemiareperfusion injury. Key words: A-kinase anchoring protein 5 (AKAP5), protein kinase A (PKA), sarcoendoplasmic reticulum calcium transport ATPase (SERCA), hypoxia and reoxygenation (H/R), apoptosis. La proteine AKAP5 (A-kinase anchoring protein 5) possede des activites biologiques variees. Cette etude a examine si AKAP5 est impliquee dans l'apoptose des cardiomyocytes induite par l'hypoxie/reoxygenation (H/R) ainsi que le mecanisme d'action sous-jacent. Des cellules H9C2 ont ete utilisees pour etablir un modele d'H/R in vitro, suivi par la surexpression d'AKAP5 dans les cellules. La cytometrie en flux a ete utilisee pour detecter le taux d'apoptose des cardiomyocytes. L'expression du phospholambane (PLN) phosphoryle, de SERCA2a et de proteines liees a l'apoptose a ete determinee par transfert Western. La coloration par immunofluorescence et l'immunoprecipitation ont ete utilisees pour detecter la distribution et l'interaction entre AKAP5, PKA et PLN. Apres l'induction de l'H/R, les cellules H9C2 montraient une expression significativement reduite de la proteine AKAP5. La regulation a la hausse d'AKAP5 favorisait la survie cellulaire et reduisait de maniere significative le niveau de LDH et le taux d'apoptose des cellules H9C2. En outre, la surexpression d'AKAP5 s'accompagnait de l'activation de la voie de signalisation PLN/SERCA2a et d'une reduction de l'apoptose. La coloration par immunofluorescence et l'immunoprecipitation ont revele que AKAP5 etait colocalisee et interagissait avec PLN et PKA. Il est interessant de noter que St-Ht31, un peptide inhibiteur qui perturbe les interactions entre AKAP et les sous-unites regulatrices, inhibait l'effet de la surexpression d'AKAP5 sur l'apoptose induite par l'H/R dans les cardiomyocytes H9C2. La surexpression d'AKAP5 attenuait l'apoptose des cardiomyocytes induite par l'H/R, possiblement par l'intermediaire de l'ancrage a la PKA pour medier la voie PLN/SERCA, ce qui suggere que AKAP5 est une cible therapeutique potentielle pour la prevention et le traitement des lesions d'ischemie-reperfusion. [Traduit par la Redaction] Mots-cles : AKAP5, proteine kinase A (PKA), ATPase de transport du calcium du SERCA (sarco/endoplasmic reticulum calcium ATPase), hypoxie et reoxygenation (H/R), apoptose., Introduction Cardiovascular diseases (CVDs) account for a large proportion of all human diseases. The incidence and mortality of acute myocardial infarction (AMI) remains high (Boateng and Sanborn 2013; Virani et [...]

Metformin, a traditional first-line pharmacological treatment for type 2 diabetes, has recently been shown to have anti-cancer effects on hepatocellular carcinoma (HCC). However, the molecular mechanism underlying the anti-tumor activity of metformin remains unclear. The Sonic hedgehog (Shh) signaling pathway is closely associated with the initiation and progression of HCC. Therefore, the aim of the current study was to investigate the effects of metformin on the biological behavior of HCC and the underlying functional mechanism of metformin in the Shh pathway. HCC was induced in HepG2 cells using recombinant human Shh (rhShh). The effects of metformin on proliferation and metastasis were evaluated using in vitro proliferation, wound healing, and invasion assays. The mRNA and protein expression levels of proteins related to the Shh pathway were measured using western blotting, quantitative PCR, and immunofluorescence staining. Metformin inhibited rhShh-induced proliferation and metastasis. Furthermore, metformin decreased the mRNA and protein expression of Shh pathway components, including Shh, Ptch, Smo, and Gli-1. Silencing of AMPK in the presence of metformin revealed that metformin exerted its inhibitory effects via AMPK. Our findings demonstrate that metformin suppresses the migration and invasion of HepG2 cells via AMPK-mediated inhibition of the Shh pathway. Key words: liver cancer, Sonic hedgehog signaling pathway, metformin, AMPK, HepG2 cells. La metformine, un traitement pharmacologique conventionnel de premiere ligne pour le diabete de type 2, s'est recemment revelee exercer des effets anticancereux sur le carcinome hepatocellulaire (CHC). Cependant, le mecanisme moleculaire de la metformine quant a son activite antitumorale n'est pas encore completement elucide. La voie de signalisation Sonic Hedgehog (Shh) est etroitement associee a l'initiation et a la progression du CHC. Ainsi, l'objectif de la presente etude consistait a examiner les effets de la metformine sur le comportement biologique du CHC et le mecanisme fonctionnel sous-jacent de la metformine sur la voie Shh. Le CHC a ete induit dans les cellules HepG2 par la Shh humaine recombinante (rhShh). Les effets de la metformine sur la proliferation et la metastase ont ete evalues par des tests de proliferation, de cicatrisation et d'invasion in vitro. Les niveaux d'expression de l'ARNm et des proteines liees a la voie Shh ont ete mesures par transfert Western, PCR quantitative et coloration par immunofluorescence. La metformine inhibait la proliferation et la metastase induites par rhShh. En outre, la metformine diminuait l'expression de l'ARNm et des proteines des composants de la voie Shh, notamment Shh, Ptch, Smo et Gli-1. Le silencage de l'AMPK en presence de metformine a revele que la metformine pouvait exercer son effet inhibiteur par l'intermediaire de l'AMPK. Ces resultats demontrent que la metformine peut supprimer la migration et l'invasion des cellules HepG2 par une inhibition de la voie Shh mediee par l'AMPK. [Traduit par la Redaction] Mots-cles : cancer du foie, voie de signalisation Sonic Hedgehog, metformine, AMPK, cellules HepG2., Introduction In 2020, liver cancer was the sixth most commonly diagnosed cancer and the third leading cause of cancer-related deaths worldwide (Sung et al. 2021). The risk factors for liver [...]

Recognition of microbe-associated molecular patterns (MAMPs) by pattern recognition receptors (PRRs) triggers the first line of inducible defence against invading pathogens.sup.1-3. Receptor-like cytoplasmic kinases (RLCKs) are convergent regulators that associate with multiple PRRs in plants.sup.4. The mechanisms that underlie the activation of RLCKs are unclear. Here we show that when MAMPs are detected, the RLCK BOTRYTIS-INDUCED KINASE 1 (BIK1) is monoubiquitinated following phosphorylation, then released from the flagellin receptor FLAGELLIN SENSING 2 (FLS2)-BRASSINOSTEROID INSENSITIVE 1-ASSOCIATED KINASE 1 (BAK1) complex, and internalized dynamically into endocytic compartments. The Arabidopsis E3 ubiquitin ligases RING-H2 FINGER A3A (RHA3A) and RHA3B mediate the monoubiquitination of BIK1, which is essential for the subsequent release of BIK1 from the FLS2-BAK1 complex and activation of immune signalling. Ligand-induced monoubiquitination and endosomal puncta of BIK1 exhibit spatial and temporal dynamics that are distinct from those of the PRR FLS2. Our study reveals the intertwined regulation of PRR-RLCK complex activation by protein phosphorylation and ubiquitination, and shows that ligand-induced monoubiquitination contributes to the release of BIK1 family RLCKs from the PRR complex and activation of PRR signalling. The detection of microorganism-associated ligands by plant cells activates a signalling cascade in which the kinase BIK1 is monoubiquinated, released from the FLS2-BAK1 complex, and internalized by endocytosis., Author(s): Xiyu Ma [sup.1] [sup.2] , Lucas A. N. Claus [sup.3] [sup.4] , Michelle E. Leslie [sup.5] [sup.10] , Kai Tao [sup.6] , Zhiping Wu [sup.7] [sup.8] , Jun Liu [...]

Mitochondria affect cellular functions alone or in cooperation with other cellular organelles. Recent research has demonstrated the close relationship of mitochondria with the endoplasmic reticulum (ER), both at the physical and the functional level. In an effort to define the combined effect of mitochondrial dysfunction (MD) and ER stress in the proinflammatory activities of macrophages, the human macrophage-like monocytic leukemia cell line THP-1 was treated with mitochondrial electron transport chain (ETC) blockers, and changes in the cellular responses upon stimulation by interferon (IFN)-[gamma] were analyzed. Inducing mitochondrial dysfunction (MD) with ETC blockers resulted in suppression of IFN-induced activation of JAK1 and STAT1/3, as well as the expression of STAT1-regulated genes. In addition, experiments utilizing pharmacological modulators of adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) and liver kinase B1 (LKBl)-deficient HeLa cells demonstrated that these suppressive effects are mediated by the LKB1-AMPK pathway. Treatment with pharmacological inhibitors of ER stress sensors failed to affect these processes, thus indicating that involvement of ER stress is not required. These results indicate that MD, induced by blocking the ETC, affects IFN-induced activation of JAK-STAT and associated inflammatory changes in THP-1 cells through the LKB1-AMPK pathway independently of ER stress. Key words: macrophage, inflammation, mitochondrial dysfunction, ER stress, JAK-STAT, AMPK. Les mitochondries affectent les fonctions cellulaires, seules ou en coopération avec d'autres organites cellulaires. La recherche récente a démontré une étroite relation entre la mitochondrie et le réticulum endoplasmique (RE), tant sur le plan physique que fonctionnel. Dans le but de définir les effets combinés de la dysfonction mitochondriale (DM) et du stress du RE sur les activités proinflammatoires des macrophages, la lignée cellulaire monocytique humaine de type macrophage THP-1 a été traitée avec des bloqueurs de la chaîne de transport d'électrons (CTE), et les changements des réponses cellulaires en réponse à une stimulation par l'interféron (IFN)-_ ont été analysés. L'induction de la dysfonction mitochondriale par les bloqueurs de la CTE donnait lieu à la suppression de l'activation de JAK1 et de STAT1/3 induite par l'IFN, de même que de l'expression des gènes régulés par STAT1. De plus, des expériences réalisées avec des modulateurs pharmacologiques de la protéine kinase activée par l'adénosine 5=-monophosphate (AMP) (AMPK) et des cellules HeLa déficientes en kinase B1 du foie (LKB1) ont démontré que ces effets suppresseurs sont médiés par la voie LKB1-AMPK. Un traitement avec des inhibiteurs pharmacologiques de détecteurs du stress du RE n'affectait pas ces processus, indiquant alors que l'implication du stress du RE n'est pas requise. Ces résultats indiquent que la dysfonction mitochondriale induite par le blocage de la CTE affecte l'activation de JAK-STAT induite par l'IFN et les changements inflammatoires associés chez les cellules THP1 par l'intermédiaire de la voie LKB1-AMPK de manière indépendante du stress du RE. [Traduit par la Rédaction] Mots-clés : macrophage, inflammation, dysfonction mitochondriale, stress du RE, JAK-STAT, AMPK., Introduction Mitochondria generate ATP by oxidative phosphorylation. This involves a redox reaction whereby electrons are transferred by the electron transport chain (ETC). Electrons are donated by nicotinamide adenine dinucleotide (NADH) [...]

The DNA-dependent protein kinase (DNA-PK), which comprises the KU heterodimer and a catalytic subunit (DNA-PKcs), is a classical non-homologous end-joining (cNHEJ) factor.sup.1. KU binds to DNA ends, initiates cNHEJ, and recruits and activates DNA-PKcs. KU also binds to RNA, but the relevance of this interaction in mammals is unclear. Here we use mouse models to show that DNA-PK has an unexpected role in the biogenesis of ribosomal RNA (rRNA) and in haematopoiesis. The expression of kinase-dead DNA-PKcs abrogates cNHEJ.sup.2. However, most mice that both expressed kinase-dead DNA-PKcs and lacked the tumour suppressor TP53 developed myeloid disease, whereas all other previously characterized mice deficient in both cNHEJ and TP53 expression succumbed to pro-B cell lymphoma.sup.3. DNA-PK autophosphorylates DNA-PKcs, which is its best characterized substrate. Blocking the phosphorylation of DNA-PKcs at the T2609 cluster, but not the S2056 cluster, led to KU-dependent defects in 18S rRNA processing, compromised global protein synthesis in haematopoietic cells and caused bone marrow failure in mice. KU drives the assembly of DNA-PKcs on a wide range of cellular RNAs, including the U3 small nucleolar RNA, which is essential for processing of 18S rRNA.sup.4. U3 activates purified DNA-PK and triggers phosphorylation of DNA-PKcs at T2609. DNA-PK, but not other cNHEJ factors, resides in nucleoli in an rRNA-dependent manner and is co-purified with the small subunit processome. Together our data show that DNA-PK has RNA-dependent, cNHEJ-independent functions during ribosome biogenesis that require the kinase activity of DNA-PKcs and its phosphorylation at the T2609 cluster. The catalytic subunit of DNA-PK autophosphorylates and contributes to ribosome biogenesis and haematopoiesis by binding to the U3 small nucleolar RNA., Author(s): Zhengping Shao [sup.1] [sup.2] , Ryan A. Flynn [sup.3] , Jennifer L. Crowe [sup.1] [sup.4] , Yimeng Zhu [sup.1] [sup.2] , Jialiang Liang [sup.5] , Wenxia Jiang [sup.1] [sup.2] [...]

The protection of telomere ends by the shelterin complex prevents DNA damage signalling and promiscuous repair at chromosome ends. Evidence suggests that the 3' single-stranded telomere end can assemble into a lasso-like t-loop configuration.sup.1,2, which has been proposed to safeguard chromosome ends from being recognized as DNA double-strand breaks.sup.2. Mechanisms must also exist to transiently disassemble t-loops to allow accurate telomere replication and to permit telomerase access to the 3' end to solve the end-replication problem. However, the regulation and physiological importance of t-loops in the protection of telomere ends remains unknown. Here we identify a CDK phosphorylation site in the shelterin subunit at Ser365 of TRF2, whose dephosphorylation in S phase by the PP6R3 phosphatase provides a narrow window during which the RTEL1 helicase can transiently access and unwind t-loops to facilitate telomere replication. Re-phosphorylation of TRF2 at Ser365 outside of S phase is required to release RTEL1 from telomeres, which not only protects t-loops from promiscuous unwinding and inappropriate activation of ATM, but also counteracts replication conflicts at DNA secondary structures that arise within telomeres and across the genome. Hence, a phospho-switch in TRF2 coordinates the assembly and disassembly of t-loops during the cell cycle, which protects telomeres from replication stress and an unscheduled DNA damage response. A phospho-switch is identified in the shelterin subunit TRF2 that regulates transient recruitment of the RTEL1 helicase to, and release from, telomeres, and provides a narrow window during which RTEL1 can unwind t-loops to facilitate telomere replication., Author(s): Grzegorz Sarek [sup.1] , Panagiotis Kotsantis [sup.1] , Phil Ruis [sup.1] , David Van Ly [sup.2] [sup.3] , Pol Margalef [sup.1] , Valerie Borel [sup.1] , Xiao-Feng Zheng [sup.4] [...]

The aspartate-specific cysteine protease caspase-8 suppresses necroptotic cell death mediated by RIPK3 and MLKL. Indeed, mice that lack caspase-8 die in a RIPK3- and MLKL-dependent manner during embryogenesis.sup.1-3. In humans, caspase-8 deficiency is associated with immunodeficiency.sup.4 or very early onset inflammatory bowel disease.sup.5. The substrates that are cleaved by caspase-8 to prevent necroptosis in vivo have not been defined. Here we show that knock-in mice that express catalytically inactive caspase-8(C362A) die as embryos owing to MLKL-dependent necroptosis, similar to caspase-8-deficient mice. Thus, caspase-8 must cleave itself, other proteins or both to inhibit necroptosis. Mice that express caspase-8(D212A/D218A/D225A/D387A), which cannot cleave itself, were viable, as were mice that express c-FLIP or CYLD proteins that had been mutated to prevent cleavage by caspase-8. By contrast, mice that express RIPK1(D325A), in which the caspase-8 cleavage site Asp325 had been mutated, died mid-gestation. Embryonic lethality was prevented by inactivation of RIPK1, loss of TNFR1, or loss of both MLKL and the caspase-8 adaptor FADD, but not by loss of MLKL alone. Thus, RIPK1(D325A) appears to trigger cell death mediated by TNF, the kinase activity of RIPK1 and FADD-caspase-8. Accordingly, dying endothelial cells that contain cleaved caspase-3 were abnormally abundant in yolk sacs of Ripk1.sup.D325A/D325A embryos. Heterozygous Ripk1.sup.D325A/+ cells and mice were viable, but were also more susceptible to TNF-induced cell death than were wild-type cells or mice. Our data show that Asp325 of RIPK1 is essential for limiting aberrant cell death in response to TNF, consistent with the idea that cleavage of RIPK1 by caspase-8 is a mechanism for dismantling death-inducing complexes. Caspase-8 suppresses apoptosis and necroptosis in embryonic mice by cleaving RIPK1., Author(s): Kim Newton [sup.1] , Katherine E. Wickliffe [sup.1] , Debra L. Dugger [sup.1] , Allie Maltzman [sup.1] , Merone Roose-Girma [sup.2] , Monika Dohse [sup.3] , László Komuves [sup.3] [...]

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., Introduction Carney complex (CNC) is a rare multiple neoplasia syndrome mainly associated with inactivating mutations of the PRKAR1A gene encoding the R1[alpha] subunit of the PKA, which results in overactive [...]