Towards system genetics analysis of head and neck squamous cell carcinoma using the mouse model,cellular platform,and clinical human data

Head and neck squamous cell cancer(HNSCC)is a leading global malignancy.Every year,More than 830000 people are diagnosed with HNSCC globally,with more than 430000 fatalities.HNSCC is a deadly diverse malignancy with many tumor locations and biological characteristics.It originates from the squamous epithelium of the oral cavity,oropharynx,nasopharynx,larynx,and hypopharynx.The most frequently impacted regions are the tongue and larynx.Previous investigations have demonstrated the critical role of host genetic susceptibility in the progression of HNSCC.Despite the advances in our knowledge,the improved survival rate of HNSCC patients over the last 40 years has been limited.Failure to identify the molecular origins of development of HNSCC and the genetic basis of the disease and its biological heterogeneity impedes the development of new therapeutic methods.These results indicate a need to identify more genetic factors underlying this complex disease,which can be better used in early detection and prevention strategies.The lack of reliable animal models to investigate the underlying molecular processes is one of the most significant barriers to understanding HNSCC tumors.In this report,we explore and discuss potential research prospects utilizing the Collaborative Cross mouse model and crossing it to mice carrying single or double knockout genes(e.g.Smad 4 and P53 genes)to identify genetic factors affecting the development of this complex disease using genome-wide association studies,epigenetics,micro RNA,long noncoding RNA,lnc RNA,histone modifications,methylation,phosphorylation,and proteomics.

Role of the metabolism of branched-chain amino acids in the development of Alzheimer’s disease and other metabolic disorders

Alzheimer’s disease is an incurable chronic neurodegenerative disorder and the leading cause of dementia,imposing a growing economic burden upon society.The disease progression is associated with gradual deposition of amyloid plaques and the formation of neurofibrillary tangles within the brain parenchyma,yet severe dementia is the culminating phase of the enduring pathology.Converging evidence suggests that Alzheimer’s disease-related cognitive decline is the outcome of an extremely complex and persistent pathophysiological process.The disease is characterized by distinctive abnormalities apparent at systemic,histological,macromolecular,and biochemical levels.Moreover,besides the well-defined and self-evident characteristic profuse neurofibrillary tangles,dystrophic neurites,and amyloid-beta deposits,the Alzheimer’s disease-associated pathology includes neuroinflammation,substantial neuronal loss,apoptosis,extensive DNA damage,considerable mitochondrial malfunction,compromised energy metabolism,and chronic oxidative stress.Likewise,distinctive metabolic dysfunction has been named a leading cause and a hallmark of Alzheimer’s disease that is apparent decades prior to disease manifestation.State-of-theart metabolomics studies demonstrate that altered branched-chain amino acids(BCAAs)metabolism accompanies Alzheimer’s disease development.Lower plasma valine levels are correlated with accelerated cognitive decline,and,conversely,an increase in valine concentration is associated with reduced risk of Alzheimer’s disease.Additionally,a clear BCAAs-related metabolic signature has been identified in subjects with obesity,diabetes,and atherosclerosis.Also,arginine metabolism is dramatically altered in Alzheimer’s disease human brains and animal models.Accordingly,a potential role of the urea cycle in the Alzheimer’s disease development has been hypothesized,and preclinical studies utilizing intervention in the urea cycle and/or BCAAs metabolism have demonstrated clinical potential.Continual failures to offer a competent treatment strategy directed against amyloid-beta or Tau proteins-related lesions,which could face all challenges of the multifaceted Alzheimer’s disease pathology,led to the hypothesis that hyperphosphorylated Tau and deposited amyloid-beta proteins are just hallmarks or epiphenomena,but not the ultimate causes of Alzheimer’s disease.Therefore,approaches targeting amyloid-beta or Tau are not adequate to cure the disease.Accordingly,the modern scientific vision of Alzheimer’s disease etiology and pathogenesis must reach beyond the hallmarks,and look for alternative strategies and areas of research.

Arginase as a Potential Target in the Treatment of Alzheimer’s Disease

Alzheimer’s disease (AD) is a slowly progressive, neurodegenerative disorder with an insidious onset that is characterized by severe decline in memory, thinking and reasoning skills. Advanced age is a prominent risk factor for AD and other metabolic diseases, such as type II diabetes and atherosclerosis. Their causal mechanisms are multifaceted and not fully understood. The precise pathophysiology of AD remains a mystery despite decades of intensive investigation. Thus far, there is no truly successful AD therapy. Arginase is the central enzyme of the urea cycle. Recent studies have identified arginase function in the brain and associated this enzyme with the development of neurodegenerative diseases. Upregulation of arginase has been shown to contribute to endothelial dysfunction, ischemia-reperfusion, atherosclerosis, diabetes, and neurodegeneration. Other state-of-the-art discoveries of the precise molecular machinery of neurodegeneration have provided new directions for the rational development of innovative therapeutic strategies in the treatment of common neurodegenerative diseases. In this context, the regulation of arginase activity appears to be a universal approach in interfering with the pathogenesis of AD and providing relief for it and other metabolic disorders. Therefore, the enzyme represents a novel therapeutic target. Arginase inhibition has been shown to reverse amyloid-driven neuronal dysfunction and microgliosis and prevent the development of other AD symptoms in rodent models of AD. Consequently, the methodology represents a promising direction for clinical development.

L-Norvaline, a new therapeutic agent against Alzheimer’s disease

Growing evidence highlights the role of arginase activity in the manifestation of Alzheimer’s disease(AD).Upregulation of arginase was shown to contribute to neurodegeneration.Regulation of arginase activity appears to be a promising approach for interfering with the pathogenesis of AD.Therefore,the enzyme represents a novel therapeutic target.In this study,we administered an arginase inhibitor,L-norvaline(250 mg/L),for 2.5 months to a triple-transgenic model(3×Tg-AD)harboring PS1M146V,APPSwe,and tauP301L transgenes.Then,the neuroprotective effects of L-norvaline were evaluated using immunohistochemistry,proteomics,and quantitative polymerase chain reaction assays.Finally,we identified the biological pathways activated by the treatment.Remarkably,L-norvaline treatment reverses the cognitive decline in AD mice.The treatment is neuroprotective as indicated by reduced beta-amyloidosis,alleviated microgliosis,and reduced tumor necrosis factor transcription levels.Moreover,elevated levels of neuroplasticity related postsynaptic density protein 95 were detected in the hippocampi of mice treated with L-norvaline.Furthermore,we disclosed several biological pathways,which were involved in cell survival and neuroplasticity and were activated by the treatment.Through these modes of action,L-norvaline has the potential to improve the symptoms of AD and even interferes with its pathogenesis.As such,L-norvaline is a promising neuroprotective molecule that might be tailored for the treatment of a range of neurodegenerative disorders.The study was approved by the Bar-Ilan University Animal Care and Use Committee(approval No.82-10-2017)on October 1,2017.

A genome-wide scan for pleiotropy between bone mineral density and nonbone phenotypes

Osteoporosis is the most common metabolic bone disorder globally and is characterized by skeletal fragility and microarchitectural deterioration.Genetic pleiotropy occurs when a single genetic element is associated with more than one phenotype.We aimed to identify pleiotropic loci associated with bone mineral density(BMD)and nonbone phenotypes in genome-wide association studies.In the discovery stage,the NHGRI-EBI Catalog was searched for genome-wide significant associations(P value<5 x 108),excluding bone-related phenotypes.SNiPA was used to identify proxies of the significantly associated single nucleotide polymorphisms(SNPs)(r2=1).We then assessed putative genetic associations of this set of SNPs with femoral neck(FN)and lumbar spine(LS)BMD data from the GEFOS Consortium.Pleiotropic variants were claimed at a false discovery rate<1.4x 103 for FN-BMD and<1.5 x 10-3 for LS-BMD.Replication of these genetic markers was performed among more than 400000 UK Biobank participants of European ancestry with available genetic and heel bone ultrasound data.In the discovery stage,72 BMD-related pleiotropic SNPs were identified,and 12 SNPs located in 11 loci on 8 chromosomes were replicated in the UK Biobank.These SNPs were associated,in addition to BMD,with 14 different phenotypes.Most pleiotropic associations were exhibited by rs479844(AP5B1,O V O L 1 genes),which was associated with dermatological and allergic diseases,and rs4072037(M U C 1 gene),which was associated with magnesium levels and gastroenterological cancer.In conclusion,12 BMD-related genome-wide significant SNPs showed pleiotropy with nonbone phenotypes.Pleiotropic associations can deepen the genetic understanding of bone-related diseases by identifying shared biological mechanisms with other diseases or traits.

Neurogenesis versus neurodegeneration:the broken balance in Alzheimer's disease

Alzheimer's disease(AD)is a progressive neurodegenerative disorder,eventually manifesting in severe cognitive dysfunction.Despite the recent proliferation of encouraging preclinical studies and clinical trials,scientific society is still far from a complete consensus regarding the AD etiology and pathogenesis.Accordingly,no approved AD-modifying therapies are currently available.Nevertheless,novel concepts predicated upon the latest discoveries and comprehension of the disease as a multifactorial disorder are paving the road to the successful AD treatment.

Baseline moderate-range albuminuria is associated with protection against severe COVID-19 pneumonia

BACKGROUND Diabetes mellitus is considered a leading contributor to severe coronavirus disease 2019(COVID-19).AIM To characterize differences between hospitalized diabetic patients with vs without COVID-19,and parameters associated with COVID-19 severity for prediction.METHODS This case-control study included 209 patients with type 2 diabetic mellitus hospitalized at the Galilee Medical Center(Nahariya,Israel)and recruited between September 2020 and May 2021,65 patients with COVID-19 infection in dedicated wards and 144 COVID-19-negative patients in internal medicine wards hospitalized due to other reasons.Clinical parameters-including age,type of antiglycemic medications,presence of retinopathy,smoking history,body mass index(BMI),glycosylated hemoglobin,maximum neutrophil:lymphocyte ratio(NLR_(max)),C-reactive protein(CRP),estimated glomerular filtration rate(eGFR),and albumin(blood and urine)-were compared between the two primary patient groups,and then between COVID-19-negative patients hospitalized due to infectious vs non-infectious disease.Finally,we explored which parameters were associated with severe COVID-19 pneumonia.RESULTS COVID-19-negative patients were older(63.9±9.9 vs 59.8±9.2,P=0.005),and had longer duration of diabetes(P=0.031),lower eGFR(P=0.033),higher albumin(P=0.026),lower CRP(P<0.001),greater smoking prevalence(P<0.001),and more baseline albuminuria(54.9%vs 30.8%,P=0.005)at admission;70%of COVID-19 patients with albuminuria had moderate-range albuminuria(albumin:creatinine 30-300 mg/g).Most of the patients with albuminuria had chronic kidney disease stage II(CKD II).Oral antiglycemic therapies were not significantly different between the two groups.Multivariable logistic regression showed that higher BMI was significantly associated with severe COVID-19(OR 1.24,95%CI:1.01-1.53,P=0.04),as was higher NLR_(max)(OR 1.2,95%CI:1.06-1.37,P=0.005).Surprisingly,pre-hospitalization albuminuria,mostly moderate-range,was associated with reduced risk(OR 0.09,95%CI:0.01-0.62,P=0.015).Moderate-range albuminuria was not associated with bacterial infections.CONCLUSION Moderate-range albuminuria in COVID-19-positive diabetic patients with CKD II is associated with less severe COVID-19.Further studies should explore this potential biomarker for risk of COVID-19-related deterioration and early interventions.

Systemic review of the robustness of randomized controlled trials for the treatment of cholangiocarcinoma in three domains: survival-inferred fragility index, restricted mean survival time, and the spin effect

Background:The vast majority of patients with cholangiocarcinoma(CC)have advanced disease at diagnosis and are candidates for palliative treatment only.The robustness of the randomized controlled trials regarding the treatment of CC are assessed.Methods:A systematic review of all randomized control trials(RCT)of treatments for both intra-and extrahepatic CC between 2010 and 2020 was performed.The survival-inferred fragility index(SIFI;the minimum number of reassignments of the best survivors between arms that would overturn the statistical outcomes)was calculated.In addition,the gain,or loss,in survival in RCTs was evaluated by the restricted mean survival time(RMST)difference.Finally,the level of spin i.e.,misrepresentation of study outcomes,was measured in inconclusive studies to assess distorted reporting strategies.Results:Out of 6,167 studies retrieved,11 could be retained for full text revision(7 with both intra-and extrahepatic CC,3 with peri-hilar CC,and 1 with peri-hilar or distal CC).Only 3 studies included resected patients(2 with both intra-and extrahepatic CC and 1 with peri-hilar or distal CC).Nine studies investigated systemic chemotherapy(including 3 after surgical resection),one study evaluated photodynamic therapy,and another investigated the use of an endoscopically inserted stent in the biliary tract.The median SIFI was−2[interquartile range(IQR):−6.25,−0.25]across all studies.Overall,the median RMST difference was 0.56 months(IQR:0.10,0.95).Finally,for inconclusive studies,the level of spin was high,moderate,and low in respectively 12.5%,25%,and 62.5% of the studies.Conclusions:RCTs of CC showed a low degree of robustness with a frequent proportion of associated spin.

The quest for optimal and reliable guidelines based on robust evidence for the treatment of cholangiocarcinoma

In 1991,Gordon H.Guyatt described the evidence-based medicine(EBM)as“a focus on educating front-line clinicians in assessing the credibility of research evidence,understanding the results of clinical studies,and determining how best to apply the results to their everyday practice”(1).In 2010,Graham et al.defined clinical guidelines as“statements that include recommendations intended to optimize patient care that are informed by a systematic review of the evidence and an assessment of the benefits and harms of alternative care options”(2).Since 2010,the number of clinical guidelines increased exponentially,and the query on PubMed with the term“new guidelines”produces 59,773 results.