The role of interaction between hepatocyte growth factor and heparan sulfate proteoglycan in the progression of hepatocellular carcinoma

Hepatosit proliferasyonunun gerçekleşmesinde önemli rolü olan sinyal ileti yolaklarından birisi de HGF-SF/c-Met sinyal yolağıdır. Birçok tümör tipinde bu yolakta bozukluklar olduğu ve bu bozuklukların hücre proliferasyonu, invazyonu ve metastazındaki artış ile sonuçlandığı bildirilmektedir. Birçok büyüme faktörü gibi HGF/SF'in biyolojik etkilerini gerçekleştirmesinde ko-reseptör gibi çalışan ve büyüme faktörlerini modüle eden Heparan Sülfat Proteoglikanların (HSPG) önemli rolü olduğu bilinmektedir. Birçok kanser tipinde heparininin HGF ile uyarılan hücre invazyonunu, hücre proliferasyonunu ve hücre hareketliliğini arttırdığı bilinmektedir. Buna karşın heparinin HCC'de hücre proliferasyonunu baskıladığı gösterilmiştir. Daha önceki çalışmamızda heparin, HGF ile uyarılan hücre proliferasyonunu, hücre invazyonunu ve hücre hareketliliğini baskıladığı belirlenmişti. Bu etkinin mekanizmasının anlaşılması için yapılan mikroarray çalışması sonrasında aday genlerden birinin Egr1 olabileceği belirlenmiştir. HCC'de HGF ile uyarılan biyolojik yanıtların düzenlenmesinde Egr-1 in rolünü incelemek amacıyla önce heparinin ve HGF uygulamasının HGF-bağımlı olan hücre invazyonuna etkisi incelendi. Egr1 ekpresyonundaki ve transkripsiyonel aktivitesindeki değişim sırası ile RT-PCR ve pGL2-luc-B-Egr1 vektörü kullanılarak lusiferaz deneyi ile gösterildi. HGF ve heparin varlığında Egr1 değişiminin hangi moleküler mekanizma ile gerçekleştiğinin belirlenmesi amacı ile gerçekleştirilen immünoblotlamalar sonrasında, heparinin, HGF ile uyarılan Egr1 aktivasyonunu, p-Met ve p-MAPK fosforilasyonu baskıladığı, ancak c-Met ve MAPK fosforile olmayan formları değişmediği saptandı. Çalışmamız sonuç olarak HGF ile uyarılan Egr1 ekspresyonunun heparin varlığında baskılanmasında MAPK yolağının önemli olduğunu ve bu sürecin hücre invazyonunun baskılanmasına neden olduğunu gösterdi. Bu bulgular HCC'nin moleküler mekanizmasının anlaşılmasında ve HCC'yi hedef alan yeni ilaçların üretiminde önemli olabilir. HGF-SF/c-Met is the most potent proliferative signaling pathway for hepatocytes and its aberrancies are very important to explain proliferative and metastatic characteristics of hepatocarcinogenesis. Additionally, it is known that Heparan Sulphate Proteoglycans (HSPGs) works as a co-receptor for HGF/SF and this interaction to modulate signaling. In particular carcinomas, heparin has been found to be increased HGF-induced cell invasion, cell motility and cell proliferation. Contradictorily, increased in HSPG expression has been reported to have negative effect on cell proliferation in HCC. We previously showed that heparin inhibits HGF-induced cell proliferation, cell invasion and cell motility and further microarray analysis was performed to determine molecular mechanism behind heparin induced HGF-SF/c-Met signaling in HCC. One of the gene that disregulate heparin and/or HGF-SF induction was Egr-1 in SKHep1. Therefore Egr-1 may contribute heparin and/or HGF-SF induced in biological consequences in HCC. First we investigated the dose dependent effects of heparin on HGF-SF induced invasion, motility and proliferation of SK-Hep1 cells with boyden chamber, invasion assay . Alteration in Egr-1 expression level and its transcriptional activity were determined by RT-PCR, and lucipherase reporter assay with pGL2-luc-B-Egr1 vector, respectively. In order to define molecular mechanisms of altered Egr-1 expression, phosphorylated and unphosphorylated forms of MAPK, levels were analyzed by western blotting. We showed that heparin inhibits HGF-induced cell invasion and also HGF- induced Egr-1 expression was found to be inhibited by heparin in SKHep1 cell line as a dose dependent manner. Moreover, it is found that heparin inhibits HGF-induced transcriptional activity of Egr-1 and HGF-induced p-MET, p-MAPK in a dose dependent manner while did not affect unphosphorylated ones. Our data concluded that heparin induced Egr-1 downregulation inhibits cell invasion and induced by HGF-SF in HCC cell lines. It may be a new aspect for the understanding molecular mechanism in HCC and good target to improve new drugs

The Value of Telemedicine for the Follow-up of Patients with New Onset Type 1 Diabetes Mellitus During COVID-19 Pandemic in Turkey: A Report of Eight Cases

The current Coronavirus disease-2019 (COVID-19) pandemic has forced health care teams to look for alternative approaches to manage a great number of children with diabetes, not only in rural but also in urban locations. The aim was to assess the provision of information about follow-up of new-onset pediatric type 1 diabetes (T1D) patients, and to investigate the integration of telemedicine into routine clinical care in the long term. The changes in coefficient of variation (CV), standard deviation and percentages of time in range (TIR), time below range (TBR) and time above range were evaluated in eight children with new-onset T1D, diagnosed during the COVID-19 pandemic. The study period was two-months of follow-up using a telemedicine system. Median follow-up time was 51 (24-66) days. Two of the patients were using low glucose suspend system and six were on multiple daily injection therapy. Target TIR values were achieved in seven patients in the last televisit and, in line with recent guidelines, a TBR <70 mg/dL (<3.9 mmol/L) (level 1 hypoglycemia) of <4% and a TBR <54 mg/dL (<3.0 mmol/L) (level 2 hypoglycemia) of <1% were achieved in all patients. Seven patients achieved a CV of <36% at their last televisit. Telemedicine as an alternative follow-up tool during unusual circumstances such pandemics, even in countries where it is not routinely used, could be beneficial to achieve optimum glycemic control in patients with new-onset T1D

MOLECULAR DIAGNOSIS IN PATIENTS WITH MONOGENIC DIABETES MELLITUS, AND DETECTION OF A NOVEL CANDIDATE GENE

Aim: We aimed to investigate molecular genetic basis of monogenic diabetes (DM) and novel responsible candidate genes with targeted Next Generation Sequencing (NGS) and Whole Exome Sequencing (WES). Methods: A hundred cases presenting with clinical findings and a family history of monogenic DM were included in the study. Molecular analysis was performed using an NGS panel including 14 genes. Following targeted NGS, WES was planned in cases in whom no variant was detected. Results: Thirty different disease-causing variants in seven different genes were detected in thirty-five (35%) cases with targeted NGS approach. Most common pathogenic variant was found in GCK gene in 25 (25%) cases. Four different variants were detected in 4 (4%) patients in ABCC8 gene. In 45 of 65 cases; WES analyses were done. A heterozygous c.2635C>T(p.Gln879Ter) variant was detected in IFIH1 gene in a patient with incidental hyperglycemia. In the segregation analysis affected mother was shown to be heterozygous for the same variant. Conclusion: Molecular etiology was determined in 35% cases with the NGS targeted panel. Seventeen novel variants in monogenic DM genes have been identified. A candidate gene determined by WES analysis in a case that could not be diagnosed with NGS panel in this study