Renal ADAM10 and 17: Their Physiological and Medical Meanings

A disintegrin and metalloproteinases (ADAMs) are a Zn2+-dependent transmembrane and secreted metalloprotease superfamily, so-called “molecular scissors,” and they consist of an N-terminal signal sequence, a prodomain, zinc-binding metalloprotease domain, disintegrin domain, cysteine-rich domain, transmembrane domain and cytoplasmic tail. ADAMs perform proteolytic processing of the ectodomains of diverse transmembrane molecules into bioactive mediators. This review summarizes on their most well-known members, ADAM10 and 17, focusing on the kidneys. ADAM10 is expressed in renal tubular cells and affects the expression of specific brush border genes, and its activation is involved in some renal diseases. ADAM17 is weakly expressed in normal kidneys, but its expression is markedly induced in the tubules, capillaries, glomeruli, and mesangium, and it is involved in interstitial fibrosis and tubular atrophy. So far, the various substrates have been identified in the kidneys. Shedding fragments become released ligands, such as Notch and EGFR ligands, and act as the chemoattractant factors including CXCL16. Their ectodomain shedding is closely correlated with pathological factors, which include inflammation, interstitial fibrosis, and renal injury. Also, the substrates of both ADAMs contain the molecules that play important roles at the plasma membrane, such as meaprin, E-cadherin, Klotho, and CADM1. By being released into urine, the shedding products could be useful for biomarkers of renal diseases, but ADAM10 and 17 per se are also notable as biomarkers. Furthermore, ADAM10 and/or 17 inhibitions based on various strategies such as small molecules, antibodies, and their recombinant prodomains are valuable, because they potentially protect renal tissues and promote renal regeneration. Although temporal and spatial regulations of inhibitors are problems to be solved, their inhibitors could be useful for renal diseases

The effects of a Jewish primary school education in England on the religious observance and practice of less or non-observant parents of the pupils

The main aim of this research is to determine whether or not there has been any noticeable change in the level of religious observance and practice of less or non-observant parents which directly or indirectly can be attributed to the influence of their children and the Jewish primary school they attend. There is a frequently voiced assumption amongst those involved in Jewish education that parents, whose children attend a Jewish primary school, have increased their level of observance due to the influence of their children and the school. However, no previous research has been carried out in the United Kingdom in order to examine the basis of this premise. The purpose ofmy own research is to test this assumption in a thorough and rigorous manner by means of both questionnaires and in-depth interviews with parents of pupils attending three Jewish primary schools in England. In addition, there are two further specific areas that will be investigated as supplementary parts ofthe main research: [i] To compare the extent of similarities and differences of any such changes in religious observance between those Jewish families in England who formed part ofmy study, and those in the USA whose children attend Jewish day schools, who have also been the subject of separate research in the USA. [ii] To determine whether within the data of this research study, there is any correlation with previous research in the field of social psychology regarding causes and effects of social conformity and deviation. The data from this specific area of research will be used to focus on the effects of a crucial inter-connection between parents, children and the school. The thesis includes an examination of previous allied research and its implications relating to the nature of religious identity and changes in parental behaviour attributed to the influence of their children's Jewish education. It also contains chapters outlining the historical and social background which led to a weakening ofJewish religious observance in the UK during the zo" century and a study of the changing role of the traditional Jewish family and its effect on the levels of religious observance in Anglo-Jewry. The data from questionnaires and interviews are analysed in a thorough manner. The results and conclusions of this thesis should be of benefit to those planning and administering Jewish primary schools in the UK

Somatic mutations of CADM1 in aldosterone-producing adenomas and gap junction-dependent regulation of aldosterone production

Aldosterone-producing adenomas (APAs) are the commonest curable cause of hypertension. Most have gain-of-function somatic mutations of ion channels or transporters. Herein we report the discovery, replication and phenotype of mutations in the neuronal cell adhesion gene CADM1. Independent whole exome sequencing of 40 and 81 APAs found intramembranous p.Val380Asp or p.Gly379Asp variants in two patients whose hypertension and periodic primary aldosteronism were cured by adrenalectomy. Replication identified two more APAs with each variant (total, n = 6). The most upregulated gene (10- to 25-fold) in human adrenocortical H295R cells transduced with the mutations (compared to wildtype) was CYP11B2 (aldosterone synthase), and biological rhythms were the most differentially expressed process. CADM1 knockdown or mutation inhibited gap junction (GJ)-permeable dye transfer. GJ blockade by Gap27 increased CYP11B2 similarly to CADM1 mutation. Human adrenal zona glomerulosa (ZG) expression of GJA1 (the main GJ protein) was patchy, and annular GJs (sequelae of GJ communication) were less prominent in CYP11B2-positive micronodules than adjacent ZG. Somatic mutations of CADM1 cause reversible hypertension and reveal a role for GJ communication in suppressing physiological aldosterone production

接着分子CADM1が基軸となる慢性腎臓病の尿細管間質病変形成

慢性腎臓病では尿細管間質病変の重篤度が残腎機能を規定するが、腎生検以外にその重篤度を知るのは難しい。慢性腎臓病では尿細管上皮のIgCAM型接着分子cell adhesion molecule 1（CADM1）の細胞外切断（shedding）が亢進しており、上皮アポトーシスを誘導する。shedding産物である細胞外断片NTFに注目し、尿中CADM1濃度を測定するサンドイッチELISAを樹立した。尿細管間質病変の重篤度と糸球体濾過率とは、尿中NTF濃度が高ければ高い程、非常に強く逆相関した。従って、尿中CADM1濃度が高い時は糸球体濾過率から尿細管傷害の重篤度を推測できると判明した。In chronic kidney disease (CKD), tubulointerstitial damage correlates with progressive decline in renal function, but it is difficult to monitor the severity without renal biopsy. Tumor suppressor CADM1/TSLC1, an IgCAM-type adhesion molecule, is expressed on renal tubular cells, and its increased ectodomain shedding is suggested to contribute to tubular degeneration. A sandwich ELISA for urinary CADM1 was developed using two anti-ectodomain antibodies. Urinary CADM1 concentrations in patients with CKD were measured. Renal biopsy specimens of all patients were pathologically scored for tubulointerstitial lesions using epithelial degeneration, interstitial inflammation, and fibrosis. There was a weak inverse correlation between pathological scores and elevated GFR (eGFR). Notably, this correlation gradually increased in patients with increasing CADM1 concentrations. CADM1 appeared to be a useful marker indicating tubulointerstitial damage from eGFR levels in CKD.研究分野：実験病

Tubulointerstitial lesion formation in chronic kidney disease mediated by renal adhesion molecule CADM1

慢性腎臓病では尿細管間質病変の重篤度が残腎機能を規定するが、腎生検以外にその重篤度を知るのは難しい。慢性腎臓病では尿細管上皮のIgCAM型接着分子cell adhesion molecule 1（CADM1）の細胞外切断（shedding）が亢進しており、上皮アポトーシスを誘導する。shedding産物である細胞外断片NTFに注目し、尿中CADM1濃度を測定するサンドイッチELISAを樹立した。尿細管間質病変の重篤度と糸球体濾過率とは、尿中NTF濃度が高ければ高い程、非常に強く逆相関した。従って、尿中CADM1濃度が高い時は糸球体濾過率から尿細管傷害の重篤度を推測できると判明した。In chronic kidney disease (CKD), tubulointerstitial damage correlates with progressive decline in renal function, but it is difficult to monitor the severity without renal biopsy. Tumor suppressor CADM1/TSLC1, an IgCAM-type adhesion molecule, is expressed on renal tubular cells, and its increased ectodomain shedding is suggested to contribute to tubular degeneration. A sandwich ELISA for urinary CADM1 was developed using two anti-ectodomain antibodies. Urinary CADM1 concentrations in patients with CKD were measured. Renal biopsy specimens of all patients were pathologically scored for tubulointerstitial lesions using epithelial degeneration, interstitial inflammation, and fibrosis. There was a weak inverse correlation between pathological scores and elevated GFR (eGFR). Notably, this correlation gradually increased in patients with increasing CADM1 concentrations. CADM1 appeared to be a useful marker indicating tubulointerstitial damage from eGFR levels in CKD.研究分野：実験病

Cell Adhesion Molecule 1 Contributes to Cell Survival in Crowded Epithelial Monolayers

When epithelial cells in vivo are stimulated to proliferate, they crowd and often grow in height. These processes are likely to implicate dynamic interactions among lateral membranous proteins, such as cell adhesion molecule 1 (CADM1), an immunoglobulin superfamily member. Pulmonary epithelial cell lines that express CADM1, named NCI-H441 and RLE-6TN, were grown to become overconfluent in the polarized 2D culture system, and were examined for the expression of CADM1. Western analyses showed that the CADM1 expression levels increased gradually up to 3 times in a cell density-dependent manner. Confocal microscopic observations revealed dense immunostaining for CADM1 on the lateral membrane. In the overconfluent monolayers, CADM1 knockdown was achieved by two methods using CADM1-targeting siRNA and an anti-CADM1 neutralizing antibody. Antibody treatment experiments were also done on 6 other epithelial cell lines expressing CADM1. The CADM1 expression levels were reduced roughly by half, in association with cell height decrease by half in 3 lines. TUNEL assays revealed that the CADM1 knockdown increased the proportion of TUNEL-positive apoptotic cells approximately 10 folds. Increased expression of CADM1 appeared to contribute to cell survival in crowded epithelial monolayers

[Reviews] Heterotypic cell-cell interaction mediated by Cell Adhesion Molecule-1, CADM1: Physiological, pathological and physical aspects

[Abstract] There are a plethora of important biological events that are regulated by cellular interactions among heterotypic cell types. The most typical example is innervation to endocrine cells and skeletal and smooth muscles. Recent biological achievements have identified many molecules that control these heterotypic cell-cell interactions. Although our understandings on these events have lately made remarkable advances at molecular levels, physical aspects of cellular adhesion have not been fully examined yet. Cell Adhesion Molecule-1, CADM1, is a member of the immunoglobulin superfamily, and has been so far characterized by several independent research groups, by virtue of its multiple functions involved in tumor suppression, synaptogenesis, and spermatogenesis. CADM1plays a key role as not only simple glue among cells, but also a conductor or promoter of heterotypic intercellular communications, for example, between nerve and mast cells, and smooth muscle and mast cells. Interestingly, it is now being revealed that the efficiency of CADM1-mediated intercellular communication is closely correlated with the kinetic strength of CADM1-mediated intercellular adhesion, by applying the latest laser techniqu

Progression of Pulmonary Emphysema and Continued Increase in Ectodomain Shedding of Cell Adhesion Molecule 1 After Cessation of Cigarette Smoke Exposure in Mice

Pulmonary emphysema usually arises in cigarette smokers, and often progresses after smoking cessation and even in ex-smokers. Lung-epithelial cell adhesion molecule 1 (CADM1), an immunoglobulin superfamily member, is extracellularly shed to produce a proapoptotic C-terminal fragment (CTF) within the cell and contribute to the development of emphysema. Here, we made an ex-smoker model using C57BL/6 mice; mice (6-week-old; 5 mice per group) were exposed to passive smoke of eight cigarettes twice a day 5 days a week until 18 weeks of age, and were then left untreated until 30 weeks of age. We calculated the mean linear intercept (Lm) and the alveolar septal thickness in the lung histologic sections to estimate the alveolar space dilatation. At 18 weeks of age, Lm was marginally enlarged (P = 0.023) with a marked increase in the septal thickness (P < 0.001) in comparison with age-matched control mice (5 mice per group), while at 30 weeks, the increase in Lm was much more prominent (P = 0.006) and the septal thickness was normalized, suggesting that emphysema progressed with septal remodeling during smoking cessation. Western blot analyses of the lungs were performed for CADM1, a possible CADM1 sheddase ADAM10, an epithelial marker pan-cytokeratin, and a myofibroblastic marker α-smooth muscle actin to estimate the expression levels of CTF and ADAM10 per epithelial cell and the levels of pan-cytokeratin and αSMA per tissue. CADM1 shedding was increased in the treated mice than in control mice at both ages, in association with an increase in the CTF level at 30 weeks (P = 0.021). In total of the treated and control mice of 30 weeks of age, Lm was positively correlated with the CTF and ADAM10 levels, and pan-cytokeratin was negatively correlated with CTF, suggesting an involvement of CADM1 shedding in emphysema progression. Positive correlations were also found between CTF and ADAM10, and between ADAM10 and αSMA, suggesting that increased septal myofibroblasts might be involved in increased CADM1 shedding. Taken together, persisting increase in ectodomain shedding of CADM1 appeared to contribute to the progression of emphysema in ex-smokers, and might be accounted for by alveolar septal remodeling