Wilms' Tumor 1 and Dax-1 Modulate the Orphan Nuclear Receptor SF-1 in Sex-Specific Gene Expression

AbstractProducts of steroidogenic factor 1 (SF-1) and Wilms' tumor 1 (WT1) genes are essential for mammalian gonadogenesis prior to sexual differentiation. In males, SF-1 participates in sexual development by regulating expression of the polypeptide hormone Müllerian inhibiting substance (MIS). Here, we show that WT1 −KTS isoforms associate and synergize with SF-1 to promote MIS expression. In contrast, WT1 missense mutations, associated with male pseudohermaphroditism in Denys-Drash syndrome, fail to synergize with SF-1. Additionally, the X-linked, candidate dosage-sensitive sex-reversal gene, Dax-1, antagonizes synergy between SF-1 and WT1, most likely through a direct interaction with SF-1. We propose that WT1 and Dax-1 functionally oppose each other in testis development by modulating SF-1–mediated transactivation

がん促進性線維芽細胞由来エクソソームを標的とした前立腺がん新規診断法の開発

application/pdf進行性前立腺がんに対するホルモン療法を開始する前に、ホルモン療法が有効な患者か、それとも抗がん剤治療を早期に始めるべきか、と判断する指標を、がん細胞周囲に存在する多様な線維芽細胞から見出すことを試みた。今回、我々が同定した線維芽細胞が産生・分泌するエクソソーム内包miR-3121-3pは、前立腺がん細胞におけるがん抑制遺伝子NKX3-1遺伝子発現を増加させる鍵分子の１つである可能性を示唆した。つまり、前立腺がん微小環境には前立腺がん細胞分化を維持するよう働く線維芽細胞が存在していることから、ホルモン療法や抗がん剤治療によってがん抑制的に働く線維芽細胞が傷害されないよう守る必要がある。The tumor stroma of prostate cancer (PCa) contains functionally different populations of cancer-associated fibroblasts (CAFs). We hypothesize that cancer progression may be predicted by characterizing the CAFs. We investigated the effects of normal PrSC and 3 PCa patient-derived CAFs on cancer-related gene expressions in 4 human PCa sublines differing in androgen receptor (AR) dependency. The mRNA expression of tumor suppressor gene NKX3-1 in PCa cells having AR dependency was significantly increased by treatment with conditioned medium of PrSC and M5. Microarray profiling of exosomal miRNAs derived from PrSC and M5 identified miR- 449c-3p and miR-3121-3p for miRNAs related to NKX3-1 mRNA. The mRNA expression of NKX3-1 in PCa cells having AR dependency was increased by transfection of miRNA mimic (has-miR-3121-3p). These results suggest that fibroblast-derived miR-3121-3p may be a candidate of stromal morphogen for maintaining PCa cell differentiation in PCa cells having AR dependency.2019年度～2021年度科学研究費補助金(基盤研究(C))研究成果報告書19K0968

IL-17A Is the Critical Cytokine for Liver and Spleen Amyloidosis in Inflammatory Skin Disease

Systemic amyloidosis is recognized as a serious complication of rheumatoid arthritis or inflammatory bowel disease, but also of inflammatory skin disease. However, the detailed molecular mechanism of amyloidosis associated with cutaneous inflammation remains unclear, and therapeutic approaches are limited. Here, we investigated the pathophysiology of amyloidosis secondary to cutaneous inflammation and the therapeutic effects of Janus kinase (JAK) inhibitors by examining a mouse model of spontaneous dermatitis (KCASP1Tg mice). Moreover, KCASP1Tg mice were crossed with interleukin-17A (IL-17A) knockout mice to generate IL-17A-/KCASP1Tg and examine the role of IL-17A in amyloidosis under cutaneous inflammation. KCASP1Tg mice showed severe amyloid deposition in the liver and spleen. Increased serum-neutral fat levels and decreased lymphocyte production were observed in the spleen. Overproduction of amyloidosis was partially ameliorated by the administration of JAK inhibitors and was further improved in IL-17A-/KCASP1Tg mice. IL-17A-producing cells included CD4, gamma delta, and CD8 T cells. In summary, our results from the analysis of a mouse model of dermatitis revealed that skin-derived inflammatory cytokines can induce amyloid deposition in the liver and spleen, and that the administration of JAK inhibitors and, even more, IL-17A ablation, reduced amyloidosis. This study demonstrates that active control of skin inflammation is essential to prevent internal organ amyloidosis

A new detection system for serum fragmented cytokeratin 18 as a biomarker reflecting histologic activities of human nonalcoholic steatohepatitis

Abstract Caspase‐generated fragmented cytokeratin 18 (fCK18) is recognized as a useful noninvasive biomarker in the diagnosis of nonalcoholic fatty liver disease (NAFLD), particularly nonalcoholic steatohepatitis (NASH). However, fCK18 measurement is not applied clinically due to widely variable cut‐off values under the current enzyme‐linked immunosorbent assay platform. Therefore, we developed a highly sensitive chemiluminescent enzyme immunoassay using newly developed monoclonal antibodies against fCK18 and investigated its relevance in NASH diagnosis. Serum fCK18 levels were measured in the derivation and validation cohort. The correlation between serum fCK18 levels and NAFLD activity score (NAS), fibrosis stage, and liver function was examined. Serum fCK18 levels were significantly correlated with alanine aminotransferase (ALT), aspartate aminotransferase (AST), and gamma‐glutamyl transpeptidase. Serum fCK18 levels were significantly associated with NAS, Brunt's grade/stage, Matteoni's classification, portal inflammation, and fat accumulation in the liver. Notably, hepatocyte ballooning was the only independent variable significantly associated with serum fCK18 in the multivariate linear regression analysis. Serum fCK18 levels were significantly elevated in patients with NAFLD and nonalcoholic fatty liver (NAFL) compared to healthy individuals. They were also significantly elevated in patients with NAFL compared to NASH defined by NAS or Matteoni's classification, with area under the curve values being 0.961 (NAFLD vs. healthy), 0.913 (NAFL vs. healthy), 0.763 (NASH vs. NAFL), and 0.796 (NASH type 3–4 vs. NAFL type 1–2). These results were confirmed by a validation cohort. Notably, changes over time in serum fCK18 levels were significantly correlated with changes in ALT, AST, and the fibrosis‐4 index in 25 patients who underwent lifestyle modification. Serum fCK18 levels were significantly correlated with liver damage associated with NASH pathology. Serum fCK18 levels are accurate in distinguishing patients with NAFL or NASH from healthy individuals and may be useful to monitor NASH over time

前立腺癌細胞分化の揺らぎによる去勢抵抗性獲得機構

application/pdf去勢抵抗性前立腺癌への進展機構としてホルモン療法後に生じる癌間質構造の多様性に着目した。癌間質を構成するactiveな線維芽細胞は細胞増殖因子やサイトカイン、細胞外マトリックスの構築などを介し、前立腺癌の悪性・進展を促進させる。ひと前立腺癌患者由来線維芽細胞とヒト前立腺癌細胞をin vitro共培養したところ、いくつかの線維芽細胞は癌細胞における癌抑制遺伝子の発現量を有意に低下させた。一方で、癌遺伝子の発現量を有意に増加させる線維芽細胞も確認された。つまり、activeな線維芽細胞は癌細胞の増殖に関わるだけでなく、癌細胞の分化状態をも変化させ、去勢抵抗性の獲得に寄与している可能性が示唆された。In the tumor microenviroment, heterogeneous stromal component of prostate cancer (PCa) tissue contains multiple populations of fibroblasts that are associated with tumorigenesis. Fibroblasts secret a number of growth factors, cytokines, ECM proteins, and miRNAs that stimulate progression of PCa cells. In this study, we investigated the effects of fibroblasts on androgen-sensitive human PCa cell line LNCaP focusing on the expression of cancer-related genes. As for tumor suppressor genes, mRNA expression of NKX3-1 in LNCaP cells was decreased by co-culturing with fibroblasts. As for oncogenes, mRNA expressions of NFKB1 and SRC in LNCap cells was increased by co-culturing with fibroblasts. Our data showed that mRNA expressions of cancer-related genes in LNCaP cells were highly disturbed by co-culturing with fibroblasts. The use of PCa patients-derived fibroblasts may allow us to investigate the characteristics of aggressive fibroblasts in the development of castration-resistant PCa.2016年度～2018年度科学研究費補助金(基盤研究(C))研究成果報告書16K1100