Interindividual Variability In Response Research Articles

Pharmacogenetics of Antipsychotic Treatment in Schizophrenia.

Antipsychotics are the mainstay treatment for schizophrenia. There is large variability between individuals in their response to antipsychotics, both in efficacy and adverse effects of treatment. While the source of interindividual variability in antipsychotic response is not completely understood, genetics is a major contributing factor. The identification of pharmacogenetic markers that predict antipsychotic efficacy and adverse reactions is a growing area of research and holds the potential to replace the current trial-and-error approach to treatment selection in schizophrenia with a personalized medicine approach.In this chapter, we provide an overview of the current state of pharmacogenetics in schizophrenia treatment. The most promising pharmacogenetic findings are presented for both antipsychotic response and commonly studied adverse reactions. The application of pharmacogenetics to schizophrenia treatment is discussed, with an emphasis on the clinical utility of pharmacogenetic testing and directions for future research.

A review of literature on the pharmacogenomics of single-nucleotide polymorphisms

Pharmacogenomics is the study of how our genetic structure determines the response to a therapeutic intervention. It is a rapidly growing field that aims to elaborate the genetic basis for differences in drug response between different individuals and to use such genetic information to predict the safety, toxicity, and/or efficacy of drugs in individual patients or groups of patients. Although drug–drug interactions and environmental factors significantly contribute to interindividual variability in drug response, genetic factors (e.g., inherited variability of drug targets, drug-metabolizing enzymes, and/or drug transporters) also appear to have a major impact on drug response and disposition. When a gene variant is associated with a particular drug response in a patient, there is the potential for making clinical decisions based on genetics by adjusting the dosage or choosing a different drug. Single-nucleotide polymorphisms (SNPs), also called SNPs, are the most common type of genetic variation among people. They are basically a substitution of a single nucleotide that occurs at a specific position in the genome. They occur normally throughout a person's DNA. They occur almost once in every 1000 nucleotides on average, which means there are roughly 4–5 million SNPs in a person's genome. Most SNPs have no effect on health or development. Some of them, however, have proven to be very important in the study of human health, especially in the field of pharmacogenomics. Researchers have found SNPs that may help predict an individual's response to certain drugs, susceptibility to environmental factors such as toxins, and risk of developing particular diseases. Scientists assess gene variants affecting an individual's drug response the same way they assess gene variants associated with diseases: by identifying genetic loci associated with known drug responses, and then testing individuals whose response is unknown. SNPs account for the most common genetic differences from person to person and pharmacogenomics explores how such changes in genetic makeup effect drug responses, resistance, etc., With our literature review, we aim to study some of the effects of these SNPs on drug responses in patients. The purpose of the study was to understand and implicate the effects of SNPs in modern medicine and how it can be applied to personalize health care for every patient. A systematic literature search was carried in search of studies pertaining to personalized medicine and implications of SNPs. A thorough search through PubMed, Google Scholar, and ProQuest revealed 61 relevant studies. All study types were considered eligible. Over the past 10 years, tremendous progress has been made in cataloging human sequence variations since this high-density map can offer the required tools to develop genetically based diagnostic and therapeutic tests. When additional functional polymorphisms are known, it may be attainable to develop helpful genetic markers also as personalized medicines. In the future, the main aim is to use SNPs not only to find certain aspects to be used in the future (individualized drug therapy, development of genome-based diets, etc.) but to also unveil the details of genome evolution. A number of challenges exist today in realizing the value of a high-density map of anonymous SNPs for pharmacogenomics. Concerns about the high price of genotyping are being addressed; however, it may be several years before the price of genotyping large populations is acceptable. In addition, availability of large patient populations will be crucial for discovering and validating SNPs.

Pharmacogenomics of mycophenolic acid in kidney transplantation: Contribution of immune response-related genes

Mycophenolic acid (MPA) inhibits IMPDH, involved in the guanosine nucleotides synthesis, and prevents DNA replication in immune cells. The repression of cell and humoral immunity by MPA induces allograft tolerance preventing acute rejection in solid organ transplantation. MPA is an effective and safe drug, but genetic and non-genetic factors have been implicated in the interindividual variability of drug response. Several studies have shown the impact of variants of pharmacokinetics or pharmacodynamics-related genes on MPA response in kidney transplantation. This review explored further the influence of genes involved in the immune response on clinical outcomes of kidney recipients on short- or long-term MPA treatment. Variants in genes related to T cell activation (CD28, CTL4, ICOS, PDPC1), pro-inflammatory cytokines (IL2, IL6, IL12A, IL12B, TNF, IFNG), immunomodulatory cytokines (IL4, IL10, TGFB1), and innate immune response (CD14, TLR2, TLR4) were shown to be associated with increased risk of acute rejection, graft function or survival, chronic graft nephropathy, viral infections or MPA-induced myelotoxicity. Some of the significant pharmacogenetic associations were confirmed by meta-analyses of kidney transplantation. These findings are suggestive that variants in immune response-related genes contribute to the variability of MPA response, and have potential application as biomarkers of acute rejection in kidney transplantation.

Nutrition and Microbiome.

The prevalence of overweight and obesity has reached epidemic proportions globally over the past few decades. The search for new management approaches continues and among them, targeting the gut microbiota can be envisioned. To date, numerous data showed the involvement of the gut microbes in the regulation and control of host metabolism. There are also increasing evidences highlighting the interactions between environmental factors, intrinsic factors, gut microbiota, and metabolic diseases. Diet emerges as the most relevant factor influencing the gut microbiome. Eating habits, as well as short-term consumption of specific diets, alter the gut microbiota composition. Moreover, nutritional disorders are associated with changes of the gut microbiota composition and/or function, as shown in obesity or type 2 diabetic patients versus healthy lean subjects. Targeting the gut microbiota for improving metabolic health appears as a new approach to manage obesity and cardio-metabolic risk. In this review, we have detailed the results of human interventions targeting the gut microbiome by prebiotic supplementation, prebiotics being defined as "substrates that are selectively utilized by the host microorganisms conferring a health benefit." If the potential benefit of this approach is obvious in preclinical models, the efficacy of prebiotics in humans is less reproducible. The inter-individual variability of response to dietary intervention can be dependent on the gut microbiota and we summarized the basal gut microbiota characteristics driving the metabolic response to dieting, prebiotic and dietary fiber intervention in the context of obesity and related metabolic diseases.

Experimental-analytical approach to assessing mechanosensitive cartilage blood marker kinetics in healthy adults: dose-response relationship and interrelationship of nine candidate markers

Purpose: To determine the suitability of selected blood biomarkers of articular cartilage as mechanosensitive markers and to investigate the dose-response relationship between ambulatory load magnitude and marker kinetics in response to load. Methods: Serum samples were collected from 24 healthy volunteers before and at three time points after a 30-minute walking stress test performed on three test days. In each experimental session, one of three ambulatory loads was applied: 100% body weight (BW); 80%BW; 120%BW. Serum concentrations of COMP, MMP-3, MMP-9, ADAMTS-4, PRG-4, CPII, C2C and IL-6 were assessed using commercial enzyme-linked immunosorbent assays. A two-stage analytical approach was used to determine the suitability of a biomarker by testing the response to the stress test (criterion I) and the dose-response relationship between ambulatory load magnitude and biomarker kinetics (criterion II). Results. COMP, MMP-3 and IL-6 at all three time points after, MMP-9 at 30 and 60 minutes after, and ADAMTS-4 and CPII at immediately after the stress test showed an average response to load or an inter-individual variation in response to load of up to 25% of pre-test levels. The relation to load magnitude on average or an inter-individual variation in this relationship was up to 8% from load level to load level. There was a positive correlation for the slopes of the change-load relationship between COMP and MMP-3, and a negative correlation for the slopes between COMP, MMP-3 and IL-6 with MMP-9, and COMP with IL6. Conclusions: COMP, MMP-3, IL-6, MMP-9, and ADAMTS-4 warrant further investigation in the context of articular cartilage mechanosensitivity and its role in joint degeneration and OA. While COMP seems to be able to reflect a rapid response, MMP-3 seems to reflect a slightly longer lasting, but probably also more distinct response. MMP-3 showed also the strongest association with the magnitude of load.

Alzheimer’s Disease Therapy, and the Effects of Genetic Polymorphisms on Its Efficacy and Safety

Alzheimer’s disease (AD) continues to be a significant health burden worldwide as the global population is getting older and health care costs are escalating. AD is a heterogeneous condition having diverse phenotypes and genotypes, which is a major challenge in understanding disease etiopathogenesis. The difficulty in understanding the intricacies of the disease has led to recurrent failure of therapeutic agents in clinical trials resulting in an extremely slow and low success in the new drug discovery process for the AD. Currently approved agents for AD therapy are acetyl cholinesterase inhibitors (AchEIs) (donepezil, galantamine and rivastigmine) and n-methyl d-aspartate (NMDA) antagonist (memantine) that provide symptomatic relief only. However, extensive inter-individual variability in drug responsiveness is observed. ‘Pharmacogenomics’ which refers to how the genome of a patient might affect the treatment response to a drug, appears to play an important role in this inter-individual variability. By bringing pharmacogenomics profile of patients on AD therapy into consideration, it might be possible to gain maximum benefits from available treatments in terms of safety, therapeutic optimization and minimizing adverse effects. The purpose of this review is to provide better understanding of AD pathogenesis, challenges of current AD therapy and insight into the role of genetic polymorphism in drug response with focus on available therapeutic options in AD.

Tools in Pharmacogenomics Biomarker Identification for Cancer Patients.

The understanding of the biological differences which underlie the inter-individual variability in drug response improved the efficacy of cancer therapy in the era of precision medicine. In fact molecularly targeted drugs and immunotherapy represent a revolution in cancer treatment. The identification of genetic predictive and/or prognostic biomarkers linked to drug pharmacokinetics (PK) and pharmacodynamics (PD) is allowed by the development of high-throughput omics tools for detecting and understanding biological differences among individuals, in order to improve drug efficacy and minimize risk of toxicity. Personalized medicine in cancer treatment reduces costs of the healthcare system. Unfortunately, pharmacogenomics biomarkers discovery is influenced by complexity, need of high-quality evidence, and a validation process for regulatory purposes. This chapter is focused on the critic analysis of presently available pharmacogenomics tools for discovering or testing genetic polymorphic variants in drug metabolizing enzyme to be introduced in clinical practice for the prospective stratification of cancer patients.

The Future of Personalized Medicine in Space: From Observations to Countermeasures.

The aim of personalized medicine is to detach from a “one-size fits all approach” and improve patient health by individualization to achieve the best outcomes in disease prevention, diagnosis and treatment. Technological advances in sequencing, improved knowledge of omics, integration with bioinformatics and new in vitro testing formats, have enabled personalized medicine to become a reality. Individual variation in response to environmental factors can affect susceptibility to disease and response to treatments. Space travel exposes humans to environmental stressors that lead to physiological adaptations, from altered cell behavior to abnormal tissue responses, including immune system impairment. In the context of human space flight research, human health studies have shown a significant inter-individual variability in response to space analogue conditions. A substantial degree of variability has been noticed in response to medications (from both an efficacy and toxicity perspective) as well as in susceptibility to damage from radiation exposure and in physiological changes such as loss of bone mineral density and muscle mass in response to deconditioning. At present, personalized medicine for astronauts is limited. With the advent of longer duration missions beyond low Earth orbit, it is imperative that space agencies adopt a personalized strategy for each astronaut, starting from pre-emptive personalized pre-clinical approaches through to individualized countermeasures to minimize harmful physiological changes and find targeted treatment for disease. Advances in space medicine can also be translated to terrestrial applications, and vice versa. This review places the astronaut at the center of personalized medicine, will appraise existing evidence and future preclinical tools as well as clinical, ethical and legal considerations for future space travel.

Weight Loss at First Month and Development of Tolerance as Possible Predictors of 30 mg Phentermine Efficacy at 6 Months

The efficacy of anti-obesity drugs usually does not consider the high degree of interindividual variability in responses to the drug which could affect the decision to withdraw the drug early due to ineffectiveness or to continue therapy according to specific expectations of success. The aim of this study was to analyze body weight loss in kilograms during the first month (1 mo-BWLkg) of treatment with 30 mg phentermine and development of tolerance to phentermine, on its 6-month efficacy. One hundred sixty-six subjects with obesity were individually or jointly analyzed in the study. Subjects with 1 mo-BWLkg of <1 kg, 1–3 kg, 3–5 kg, and ≥5 kg reached 6-month mean percentage body weight reductions (BWR%) of approximately 3%, 5%, 10%, and 15%, respectively. Development of late tolerance (4–6 months) to phentermine had a lower impact than early tolerance (2–3 months). Subjects with 1 mo-BWLkg < 3 kg who developed early tolerance did not achieve relevant BWR% (≥5%) at month 6, while the rest of the subgroups achieved increasing and progressive BWR%, according to their 1 mo-BWLkg range and time of onset of tolerance. The 1 mo-BWLkg and development of tolerance to phentermine could be useful to predict the expected 6-month efficacy trends in obese patients treated with 30 mg phentermine.

MRNA and small RNA gene expression changes in peripheral blood to detect internal Ra-223 exposure

Purpose Excretion analysis is the established method for detection of incorporated alpha-emitting radionuclides, but it is laborious and time consuming. We sought a simplified method in which changes in gene expression might be measured in human peripheral blood to detect incorporated radionuclides. Such an approach could be used to quickly determine internal exposure in instances of a radiological dispersal device or a radiation accident. Materials and methods We evaluated whole blood samples from five patients with castration-resistant prostate cancer and multiple bone metastases (without visceral or nodal involvement), who underwent treatment with the alpha emitting isotope Radium-223 dichloride (Ra-223, Xofigo®). Patients received about 4 MBq per cycle and, depending on survival and treatment tolerance, were followed for six months. We collected 24 blood samples approximately monthly corresponding to treatment cycle. Results Firstly, we conducted whole genome screening of mRNAs (mRNA seq) and small RNAs (small RNA seq) using next generation sequencing in one patient at eight different time points during all six cycles of Ra-223-therapy. We identified 1900 mRNAs and 972 small RNAs (222 miRNAs) that were differentially up- or down-regulated during follow-up after the first treatment with Ra-223. Overall candidate RNA species inclusion criteria were a general (≥|2|-fold) change or with peaking profiles (≥|5|-fold) at specific points in time. Next we chose 72 candidate mRNAs and 101 small RNAs (comprising 29 miRNAs) for methodologic (n = 8 samples, one patient) and independent (n = 16 samples, four patients) validation by qRT-PCR. In total, 15 mRNAs (but no small RNAs) were validated by methodologic and independent testing. However, the deregulation occurred at different time points, showing a large inter-individual variability in response among patients. Conclusions This proof of concept provides support for the applicability of gene expression measurements to detect internalized alpha-emitting radionuclides, but further work is needed with a larger sample size. While our approach has merit for internal deposition monitoring, it was complicated by the severe clinical condition of the patients we studied.

Single-Cell and Bulk RNA-Sequencing Reveal Differences in Monocyte Susceptibility to Influenza A Virus Infection Between Africans and Europeans.

There is considerable inter-individual and inter-population variability in response to viruses. The potential of monocytes to elicit type-I interferon responses has attracted attention to their role in viral infections. Here, we use single-cell RNA-sequencing to characterize the role of cellular heterogeneity in human variation of monocyte responses to influenza A virus (IAV) exposure. We show widespread inter-individual variability in the percentage of IAV-infected monocytes. Notably, individuals with high cellular susceptibility to IAV are characterized by a lower activation at basal state of an IRF/STAT-induced transcriptional network, which includes antiviral genes such as IFITM3, MX1 and OAS3. Upon IAV challenge, we find that cells escaping viral infection display increased mRNA expression of type-I interferon stimulated genes and decreased expression of ribosomal genes, relative to both infected cells and those never exposed to IAV. We also uncover a stronger resistance of CD16+ monocytes to IAV infection, together with CD16+ -specific mRNA expression of IL6 and TNF in response to IAV. Finally, using flow cytometry and bulk RNA-sequencing across 200 individuals of African and European ancestry, we observe a higher number of CD16 + monocytes and lower susceptibility to IAV infection among monocytes from individuals of African-descent. Based on these data, we hypothesize that higher basal monocyte activation, driven by environmental factors and/or weak-effect genetic variants, underlies the lower cellular susceptibility to IAV infection of individuals of African ancestry relative to those of European ancestry. Further studies are now required to investigate how such cellular differences in IAV susceptibility translate into population differences in clinical outcomes and susceptibility to severe influenza.

Abstract 10197: Cavin1, a Novel Actor in Drug Induced Long Qt

Introduction: Many drugs have been pulled from the market due to their undesirable effect in prolonging the QT interval by altering cardiac ion currents, notably I Kr generated by hERG. This cardiotoxic impact is however extremely variable between individuals and the mechanisms underlying this variability remain undiscovered. Previously, we identified a higher expression of CAVIN1, an essential protein for caveolae biogenesis, in cardiomyocytes (CM) derived from individuals with high susceptibility to develop sotalol-induced long QT. Hypothesis: CAVIN1 can modulate cardiomyocytes’ response to drugs. Methods: We used induced pluripotent stem cells-derived-CM (iCM) from 3 subjects with the highest sensitivity (HS) to sotalol (ie, those with the highest increase in QTc) and 3 others with the lowest sensitivity to sotalol (LS). Results: cardiac repolarization and I Kr were similar between HS-iCM and LS-iCM at baseline. However, HS-iCM displayed a significantly higher prolongation of the repolarization phase along with a decrease in I Kr in response to sotalol and other drugs targeting hERG (E4031, vandetanib, and clarithromycin). CAVIN1 mRNA and protein levels were increased by 2-fold in the HS-iCM. Overexpressing CAVIN1 in the LS-iCM using adenovirus induced higher sensitivity to sotalol and the other hERG blockers. In contrast, CAVIN1 knockdown using siRNA in the HS-iCM switched their phenotype to a lower sensitivity to hERG blockers. We observed that CAVIN1 overexpression in LS-iCM and HEK cells changed hERG distribution within cells with accumulation at the membrane. We applied Methyl-β-cyclodextrin, a detergent that removes caveolae from membranes, and found that caveolae depletion induced prolonged repolarization associated with a decrease in CAVIN1 and hERG sarcolemmal expression, and their accumulation in the cytoskeleton. In contrast with the LS-iCM, sotalol application in the HS-iCM promoted hERG internalization into the cytoskeleton. Conclusions: We identified CAVIN1 as a novel modulator of cardiac repolarization in response to drug by promoting hERG internalization. This represents a novel mechanism sustaining the inter-individual variability in response to drugs.

Human carboxylesterase 1A plays a predominant role in the hydrolytic activation of remdesivir in humans

Remdesivir, an intravenous nucleotide prodrug, has been approved for treating COVID-19 in hospitalized adults and pediatric patients. Upon administration, remdesivir can be readily hydrolyzed to form its active form GS-441524, while the cleavage of the carboxylic ester into GS-704277 is the first step for remdesivir activation. This study aims to assign the key enzymes responsible for remdesivir hydrolysis in humans, as well as to investigate the kinetics of remdesivir hydrolysis in various enzyme sources. The results showed that remdesivir could be hydrolyzed to form GS-704277 in human plasma and the microsomes from human liver (HLMs), lung (HLuMs) and kidney (HKMs), while the hydrolytic rate of remdesivir in HLMs was the fastest. Chemical inhibition and reaction phenotyping assays suggested that human carboxylesterase 1 (hCES1A) played a predominant role in remdesivir hydrolysis, while cathepsin A (CTSA), acetylcholinesterase (AchE) and butyrylcholinesterase (BchE) contributed to a lesser extent. Enzymatic kinetic analyses demonstrated that remdesivir hydrolysis in hCES1A (SHUTCM) and HLMs showed similar kinetic plots and much closed Km values to each other. Meanwhile, GS-704277 formation rates were strongly correlated with the CES1A activities in HLM samples from different individual donors. Further investigation revealed that simvastatin (a therapeutic agent for adjuvant treating COVID-19) strongly inhibited remdesivir hydrolysis in both recombinant hCES1A and HLMs. Collectively, our findings reveal that hCES1A plays a predominant role in remdesivir hydrolysis in humans, which are very helpful for predicting inter-individual variability in response to remdesivir and for guiding the rational use of this anti-COVID-19 agent in clinical settings.

Nutrigenomic Effect of Hydroxytyrosol in Vascular Endothelial Cells: A Transcriptomic Profile Analysis

Hydroxytyrosol (HT), a peculiar olive and olive oil phenolic antioxidant, plays a significant role in the endothelial and cardiovascular protection associated with olive oil consumption. However, studies examining the effects of HT on the whole-genome expression of endothelial cells, which are prominent targets for vasculo-protective effects of olive oil polyphenols, have been lacking. This study aims to comprehensively evaluate the genomic effects exerted by HT, at the transcriptional level, in endothelial cells under resting or proinflammatory conditions. Human umbilical vein endothelial cells (HUVECs) were treated with 10 µmol/L HT for 1 h and then stimulated with 5 ng/mL interleukin (IL)-1β for 3 h. Total RNA was extracted, and gene expression profile assessed with microarray analysis. Functional enrichment analysis and pathway analysis were performed by Ingenuity Pathways Analysis. Microarray data were validated by qRT-PCR. Fixing a significance threshold at 1.5-fold change, HT affected the expression of 708 and 599 genes, respectively, in HUVECs under resting and IL-1β-stimulated conditions; among these, 190 were common to both conditions. Unfolded protein response (UPR) and endoplasmic reticulum stress resulted from the two top canonical pathways common between HT and HT-IL-1β affected genes. IL-17F/A signaling was found in the top canonical pathways of HT modified genes under resting unstimulated conditions, whereas cardiac hypertrophy signaling was identified among the pathways affected by HT-IL-1β. The transcriptomic analysis allowed pinpointing immunological, inflammatory, proliferative, and metabolic-related pathways as the most affected by HT in endothelial cells. It also revealed previously unsuspected genes and related gene pathways affected by HT, thus broadening our knowledge of its biological properties. The unbiased identification of novel genes regulated by HT improves our understanding of mechanisms by which olive oil prevents or attenuates inflammatory diseases and identifies new genes to be enquired as potential contributors to the inter-individual variation in response to functional food consumption.

Establishing a Novel Pipeline That Combines in-Silico Prediction with in-Vitro and Ex-Vivo Validation to Discover Secondary Drug Combinations Against Relapsed and/or Refractory Multiple Myeloma

Multiple myeloma (MM) is the second-most common hematological malignancy in the US. MM is an incurable, age-dependent plasma cell neoplasm with a 5-year survival rate of less than 50%. Extensive inter-individual variation in response to standard-of-care drugs like proteasome inhibitors (PIs) and immunomodulatory drugs (IMiDs), drug resistance, and dose-limiting toxicities are critical problems for the treatment of MM. Clinical success in anti-myeloma treatment, therefore, warrants continuous development of novel combination therapy strategies with the explicit goal to improve the therapeutic efficacy by concomitantly targeting multiple signaling pathways.Previously, we have reported the development of an in-house computational pipeline called secDrug that applies greedy algorithm-based set-covering computational optimization method followed by a regularization technique to predict secondary drugs that can be repurposed as novel synergistic partners of standard-of-care drugs for the management of refractory/ resistant MM. Top among these secondary drugs (secDrugs) were the HSP90 inhibitor 17-AAG.In this study, we used 17-AAG as a proof of principle to establish a pipeline that integrates our in silico predictions with in vitro and ex vivo validation as well as multi-omics technologies to identify, validate, and characterize therapeutic agents that could be used either alone or in combination with standard-of-care drugs for the treatment of R/R MM patients (Figure 1).To screen and validate our in silico prediction results, we performed in vitro cytotoxicity assays using 17-AAG on a panel of human myeloma cell lines (HMCLs; in vitro model systems) that captures a wide range of biological and genetic heterogeneity representing the complexities encountered in clinical settings. These cell lines include HMCLs representing innate sensitive/resistance, >10 pairs of parental and clonally-derived PI- and IMiD-resistant pairs (P vs VR or LenR; representing acquired/emerging resistance/relapse), NRAS mutants which leads to the constitutive activation of oncogenic Ras signaling, and CRISPR-edited HSP90 knockdown cell line. Our results showed that 17-AAG has high synergistic activity in combination with PI in inducing apoptosis even in innate and acquired PI-resistant HMCLs and significantly reduces the effective dose of PI required to achieve IC 50 (Chou-Talalay's Dose Reduction Index or DRI 7±1.4). Moreover, 17-AAG+IMiD showed synergistic cell killing activity in clonally-derived IMiD resistant HMCL. Further, 17-AAG induced cell death was comparable with Hsp90 knockdown as evident from the cytotoxicity assay using PI and 17-AAG in combination in RPMI8226-wild type and RPMI-HSP90AA1 knocked down cell line. Notably, 17-AAG was strikingly effective against the NRAS-mutant cell line indicating an additional niche (NRas mutant myeloma) where 17-AAG could be most effective.Next, we performed RNA sequencing to elucidate the molecular mechanisms behind 17-AAG drug action, drug synergy, 17-AAG-induced cell death. Our gene expression profiling (GEP) followed by Ingenuity Pathway Analysis (IPA) analysis revealed protein ubiquitination, aryl hydrocarbon receptor signalling pathway as the top canonical pathways. 17-AAG induced apoptosis via mitochondrial mediated pathway in myeloma. 17-AAG exerts its cytotoxic effect by activating intrinsic pathway of apoptosis which we further confirmed through the increase in reactive oxygen species generation and decrease in mitochondrial membrane potential. 17-AAG was also effective in reducing the expression of hallmarks of MM such as p65/NF-kB, IRF4, c-Myc. Finally, we performed mass cytometry (CyTOF; Cytometry by time of flight) on primary bone-marrow cells (PMCs) from myeloma patients for further validation of proteomic signatures at the single-cell level. CyTOF analysis confirmed 17-AAG-induced cell death and key changes in MM-specific proteomic markers. 17-AAG treated PMCs showed elevated cleaved caspase levels and down-regulation of IRF4 and phospho-STAT3. GEP and CyTOF results were confirmed using immunoblotting assays.Together, our study demonstrates a unique pipeline for drug repositioning that has the potential to revolutionize clinical decision-making by minimizing the number of drugs required for discovering successful combination chemotherapy regimens against drug-resistant myeloma. [Display omitted] DisclosuresKumar: BMS: Consultancy, Research Funding; Abbvie: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Amgen: Consultancy, Research Funding; Takeda: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Tenebio: Research Funding; Beigene: Consultancy; Oncopeptides: Consultancy; Antengene: Consultancy, Honoraria; Carsgen: Research Funding; Janssen: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; KITE: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Merck: Research Funding; Astra-Zeneca: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Novartis: Research Funding; Roche-Genentech: Consultancy, Research Funding; Bluebird Bio: Consultancy; Celgene: Membership on an entity's Board of Directors or advisory committees, Research Funding; Adaptive: Membership on an entity's Board of Directors or advisory committees, Research Funding; Sanofi: Research Funding.

Cerebral blood flow is associated with matrix metalloproteinase levels during the early symptomatic phase of concussion.

Concussion is associated with disrupted cerebral blood flow (CBF), although there appears to be substantial inter-individual variability in CBF response. At present, the mechanisms of variable CBF response remain incompletely understood, but one potential contributor is matrix metalloproteinase (MMP) expression. In more severe forms of acquired brain injury, MMP up-regulation contributes to CBF impairments via increased blood-brain barrier permeability. A similar relationship is hypothesized for concussion, where recently concussed individuals with higher MMP levels have lower CBF. To test this hypothesis, 35 concussed athletes were assessed longitudinally at early symptomatic injury (median: 5 days post-injury) and at medical clearance (median: 24 days post-injury), along with 71 athletic controls. For all athletes, plasma MMPs were measured and arterial spin labelling was used to measure CBF. Consistent with our hypothesis, higher concentrations of MMP-2 and MMP-3 were correlated with lower global CBF. The correlations between MMPs and global CBF were also significantly diminished for concussed athletes at medical clearance and for athletic controls. These results indicate an inverse relationship between plasma MMP levels and CBF that is specific to the symptomatic phase of concussion. Analyses of regional CBF further showed that correlations with MMP levels exhibited some spatial specificity, with greatest effects in occipital, parietal and temporal lobes. These findings provide new insights into the mechanisms of post-concussion cerebrovascular dysfunction.

Interhemispheric functional connectivity of sensorimotor cortex predicts responsiveness of transcranial magnetic stimulation in older adults

Abstract Background: Theta burst stimulation (TBS) holds promise for the induction of brain plasticity. However, the effects of TBS are highly variable across individual participants, and perhaps more so in older adults. Although it’s well established that non-invasive brain stimulation, including TBS, affects not only the stimulated region but also remote areas, how brain connectivity is modulated by TBS in older adults and whether baseline connectivity mediates the TBS effect remain unclear. The purpose of the study is to identify baseline network properties of sensorimotor system that may contribute to inter-individual variability of TBS response. Methods 19 right-handed older adults underwent one excitatory intermittent TBS (iTBS) session over left primary motor cortex (M1). We acquired both motor-evoked potentials (MEPs) at 13 single-pulse stimulation intensities and resting state fMRI data immediately before and after the iTBS session. We tested whether baseline measures including active motor threshold (aMT), input-output curve parameters and interhemispheric functional connectivity (ihFC) would predict individual response pattern. Results: iTBS significantly enhanced MEPs and functional connectivity in bilateral M1, somatosensory cortex and supplementary motor area. We found that participants with a lower aMT featured a relatively lower ihFC among motor cortices at baseline. Furthermore, our study showed that the relatively lower ihFC before iTBS session was associated with a greater enhancement in MEPs. Conclusion: Age-related reduction in hemispheric asymmetry has been frequently reported. In agreement with previous studies, older adults may recruit bilateral sensorimotor networks to maintain functionality, and thus exhibit a higher level of baseline ihFC. Similarly, the higher level of baseline ihFC might serve as a compensatory mechanism for the decreased sensitivity to single-pulse stimulation as indicated by a higher aMT. In summary, our findings suggest that older adults with lower aMT as well as lower ihFC might be able to elicit iTBS induced plasticity more effectively. Keywords: Theta burst stimulation, Cortical excitability, Interhemispheric connectivity, Inter-individual variability

Training responsiveness of cardiorespiratory fitness and arterial stiffness following moderate-intensity continuous training and high-intensity interval training in adults with intellectual and developmental disabilities.

Cardiorespiratory fitness (CRF) prompts antiatherogenic adaptations in vascular function and structure. However, there is an extraordinary interindividual variability in response to a standard dose of exercise, wherein a substantial number of adults with intellectual and developmental disabilities (IDD) do not improve CRF. We (1) evaluated the effects of 12-month of moderate-intensity continuous training (MICT) on CRF and arterial stiffness and (2) tested whether an additional 3-month of high-intensity interval training (HIIT) would add to improvements in CRF responsiveness and arterial stiffness. Fifteen adults with mild-to-moderate IDD (male adults=9, 30.1±7.5years old) met 3days per week for 30min MICT for 12months, after which the incidence of CRF responsiveness was calculated (≥5.0% change in absolute peak VO2 ). Thereafter, responders and non-responders started HIIT for 3months with identical daily training load/frequency. Peak VO2 , local and regional indices of arterial stiffness were assessed prior to and after each period. Sixty per cent of the participants were non-responders following MICT, but the incidence dropped to 20% following HIIT (P=0.03). Absolute peak VO2 values reached significant difference from pre-intervention (+0.38±0.08Lmin-1 , P=0.001) only when HIIT was added. Lower limb pulse wave velocity (PWV) decreased following MICT (-0.8±1.1ms-1 , P=0.049), whereas central PWV only decreased following HIIT (-0.8±0.9ms-1 , P=0.013). Cardiorespiratory fitness responsiveness and reductions in PWV to a 12-month MICT period in adults with IDD improved following a period of HIIT programme inducing higher metabolic stress.

Impact of 27Gln\Glu Polymorphisms of β2-Adrenergic Receptor Gene on Pulmonary Function in Asthmatic Children Treated with Nebulized Salbutamol

Background: The β2-adrenergic receptor gene is a typical candidate for the study of genetic polymorphism. Many patients with asthma do not respond to β2-agonist; in addition, a wide interindividual variability in pharmacological response exists, likely because of the interaction between clinical, environmental, and genetic factors. Methods: A total of 80 children (55 boys and 25 girls) with mild-moderate acute asthma attacks, ages range between 5 and 18 years, attended the Asthma Clinic at Karbala Teaching Hospital for Children were included in this study. Spirometry (Spirolab III) was used for lung function assessment before and after treatment with nebulized salbutamol, and allele-specific polymerase chain reaction was performed to determine single nucleotide polymorphisms (SNP) of β2-adrenergic receptor gene at nucleic acid 79 (27 amino acid position). Results: The effect of treatment with nebulized salbutamol on lung function in asthmatic children having different genotypes of 27Gln\Glu SNP demonstrated that patients having GG and CG genotypes showed significant improvement (P &lt; 0.05) in forced expiratory volume in 1 s (FEV1), forced vital capacity (FVC), FEV1/FVC ratio, and FEV1% predicted after treatment, while patients having CC genotype showed significant improvement (P &lt; 0.05) in FEV1, FVC, and FEV1% predicted after treatment with nebulized salbutamol without significant improvement in FEV1/FVC ratio. Patients having CG genotype of 27Gln\Glu SNP showed highest change (increase) in FEV1 (185.38 ± 174.93 ml) and in FEV1% predicted (14.96 ± 15.45%) after treatment with nebulized salbutamol in comparison with other genotype groups although this change was not statistically significant between groups. Conclusion: Asthmatic children having CG and GG genotypes of β2-adrenergic receptor gene (27Gln\Glu) showed better improvement in lung function after treatment with nebulized salbutamol in comparison with those having CC (27Gln\Glu) genotype during mild-to-moderate acute asthma attacks.

Severity of COVID-19 Patients Predicted by Serum Sphingolipids Signature.

The reason behind the high inter-individual variability in response to SARS-CoV-2 infection and patient’s outcome is poorly understood. The present study targets the sphingolipid profile of twenty-four healthy controls and fifty-nine COVID-19 patients with different disease severity. Sera were analyzed by untargeted and targeted mass spectrometry and ELISA. Results indicated a progressive increase in dihydrosphingosine, dihydroceramides, ceramides, sphingosine, and a decrease in sphingosine-1-phosphate. These changes are associated with a serine palmitoyltransferase long chain base subunit 1 (SPTLC1) increase in relation to COVID-19 severity. Severe patients showed a decrease in sphingomyelins and a high level of acid sphingomyelinase (aSMase) that influences monosialodihexosyl ganglioside (GM3) C16:0 levels. Critical patients are characterized by high levels of dihydrosphingosine and dihydroceramide but not of glycosphingolipids. In severe and critical patients, unbalanced lipid metabolism induces lipid raft remodeling, leads to cell apoptosis and immunoescape, suggesting active sphingolipid participation in viral infection. Furthermore, results indicated that the sphingolipid and glycosphingolipid metabolic rewiring promoted by aSMase and GM3 is age-dependent but also characteristic of severe and critical patients influencing prognosis and increasing viral load. AUCs calculated from ROC curves indicated ceramides C16:0, C18:0, C24:1, sphingosine and SPTLC1 as putative biomarkers of disease evolution.