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Background: GATA4 (GATA-binding protein 4), a zinc finger–containing, DNA-binding transcription factor, is essential for normal cardiac development and homeostasis in mice and humans, and mutations in this gene have been reported in human heart defects. Defects in alternative splicing are associated with many heart diseases, yet relatively little is known about how cell type– or cell state–specific alternative splicing is achieved in the heart. Here, we show that GATA4 regulates cell type–specific splicing through direct interaction with RNA and the spliceosome in human induced pluripotent stem cell–derived cardiac progenitors. Methods: We leveraged a combination of unbiased approaches including affinity purification of GATA4 and mass spectrometry, enhanced cross-linking with immunoprecipitation, electrophoretic mobility shift assays, in vitro splicing assays, and unbiased transcriptomic analysis to uncover GATA4’s novel function as a splicing regulator in human induced pluripotent stem cell–derived cardiac progenitors. Results: We found that GATA4 interacts with many members of the spliceosome complex in human induced pluripotent stem cell–derived cardiac progenitors. Enhanced cross-linking with immunoprecipitation demonstrated that GATA4 also directly binds to a large number of mRNAs through defined RNA motifs in a sequence-specific manner. In vitro splicing assays indicated that GATA4 regulates alternative splicing through direct RNA binding, resulting in functionally distinct protein products. Correspondingly, knockdown of GATA4 in human induced pluripotent stem cell–derived cardiac progenitors resulted in differential alternative splicing of genes involved in cytoskeleton organization and calcium ion import, with functional consequences associated with the protein isoforms. Conclusions: This study shows that in addition to its well described transcriptional function, GATA4 interacts with members of the spliceosome complex and regulates cell type–specific alternative splicing via sequence-specific interactions with RNA. Several genes that have splicing regulated by GATA4 have functional consequences and many are associated with dilated cardiomyopathy, suggesting a novel role for GATA4 in achieving the necessary cardiac proteome in normal and stress-responsive conditions.

BACKGROUND: Known genetic causes of congenital heart disease (CHD) explain METHODS: We tested de novo variants from trio studies of 2649 CHD probands and their parents, as well as rare (allele frequency, − 6 ) variants from 4472 CHD probands in the Pediatric Cardiac Genetics Consortium through a combined computational and in vitro approach. RESULTS: We identified 53 de novo and 74 rare variants in CHD cases that alter splicing and thus are loss of function. Of these, 77 variants are in known dominant, recessive, and candidate CHD genes, including KMT2D and RBFOX2 . In 1 case, we confirmed the variant’s predicted impact on RNA splicing in RNA transcripts from the proband’s cardiac tissue. Two probands were found to have 2 loss-of-function variants for recessive CHD genes HECTD1 and DYNC2H1 . In addition, SpliceAI—a predictive algorithm for altered RNA splicing—has a positive predictive value of ≈93% in our cohort. CONCLUSIONS: Through assessment of RNA splicing, we identified a new loss-of-function variant within a CHD gene in 78 probands, of whom 69 (1.5%; n=4472) did not have a previously established genetic explanation for CHD. Identification of splice-altering variants improves diagnostic classification and genetic diagnoses for CHD. REGISTRATION: URL: https://clinicaltrials.gov ; Unique identifier: NCT01196182.

Background: Congenital heart disease (CHD) is highly heritable, but the power to identify inherited risk has been limited to analyses of common variants in small cohorts. Methods: We performed reimputation of 4 CHD cohorts (n=55 342) to the TOPMed reference panel (freeze 5), permitting meta-analysis of 14 784 017 variants including 6 035 962 rare variants of high imputation quality as validated by whole genome sequencing. Results: Meta-analysis identified 16 novel loci, including 12 rare variants, which displayed moderate or large effect sizes (median odds ratio, 3.02) for 4 separate CHD categories. Analyses of chromatin structure link 13 of the genome-wide significant loci to key genes in cardiac development; rs373447426 (minor allele frequency, 0.003 [odds ratio, 3.37 for Conotruncal heart disease]; P =1.49×10 −8 ) is predicted to disrupt chromatin structure for 2 nearby genes BDH1 and DLG1 involved in Conotruncal development. A lead variant rs189203952 (minor allele frequency, 0.01 [odds ratio, 2.4 for left ventricular outflow tract obstruction]; P =1.46×10 − 8 ) is predicted to disrupt the binding sites of 4 transcription factors known to participate in cardiac development in the promoter of SPAG9 . A tissue-specific model of chromatin conformation suggests that common variant rs78256848 (minor allele frequency, 0.11 [odds ratio, 1.4 for Conotruncal heart disease]; P =2.6×10 − 8 ) physically interacts with NCAM1 ( P FDR =1.86×10 − 27 ), a neural adhesion molecule acting in cardiac development. Importantly, while each individual malformation displayed substantial heritability (observed h2 ranging from 0.26 for complex malformations to 0.37 for left ventricular outflow tract obstructive disease) the risk for different CHD malformations appeared to be separate, without genetic correlation measured by linkage disequilibrium score regression or regional colocalization. Conclusions: We describe a set of rare noncoding variants conferring significant risk for individual heart malformations which are linked to genes governing cardiac development. These results illustrate that the oligogenic basis of CHD and significant heritability may be linked to rare variants outside protein-coding regions conferring substantial risk for individual categories of cardiac malformation.

Chronic inflammation and tissue fibrosis are common stress responses that worsen organ function, yet the molecular mechanisms governing their crosstalk are poorly understood. In diseased organs, stress-induced changes in gene expression fuel maladaptive cell state transitions and pathological interaction between diverse cellular compartments. Although chronic fibroblast activation worsens dysfunction of lung, liver, kidney, and heart, and exacerbates many cancers, the stress-sensing mechanisms initiating the transcriptional activation of fibroblasts are not well understood. Here, we show that conditional deletion of the transcription co-activatorBrd4inCx3cr1-positive myeloid cells ameliorates heart failure and is associated with a dramatic reduction in fibroblast activation. Analysis of single-cell chromatin accessibility and BRD4 occupancyin vivoinCx3cr1-positive cells identified a large enhancer proximal toInterleukin-1 beta (Il1b), and a series of CRISPR deletions revealed the precise stress-dependent regulatory element that controlled expression ofIl1bin disease. Secreted IL1B functioned non-cell autonomously to activate a p65/RELA-dependent enhancer near the transcription factorMEOX1, resulting in a profibrotic response in human cardiac fibroblasts.In vivo, antibody-mediated IL1B neutralization prevented stress-induced expression ofMEOX1, inhibited fibroblast activation, and improved cardiac function in heart failure. The elucidation of BRD4-dependent crosstalk between a specific immune cell subset and fibroblasts through IL1B provides new therapeutic strategies for heart disease and other disorders of chronic inflammation and maladaptive tissue remodeling.

This year, Cell Stem Cell and the International Society for Stem Cell Research (ISSCR) are celebrating their 15

The cell-adhesion proteins neuroligin-3 and neuroligin-4X (NLGN3/4X) have well described roles in synapse formation. NLGN3/4X are also expressed highly during neurodevelopment. However, the role these proteins play during this period is unknown. Here we show that NLGN3/4X localized to the leading edge of growth cones where it promoted neuritogenesis in immature human neurons. Super-resolution microscopy revealed that NLGN3/4X clustering induced growth cone enlargement and influenced actin filament organization. Critically, these morphological effects were not induced by autism spectrum disorder (ASD)-associated NLGN3/4X variants. Finally, actin regulators p21-activated kinase 1 and cofilin were found to be activated by NLGN3/4X and involved in mediating the effects of these adhesion proteins on actin filaments, growth cones and neuritogenesis. These data reveal a novel role for NLGN3 and NLGN4X in the development of neuronal architecture, which may be altered in the presence of ASD-associated variants.

The nanocomposite surface coating film samples were prepared by high-speed mechanical mixing of novolac epoxy resin (EEW 180 eq/gm) and a varying amount of nano-CaCO3 (0–5 wt%) for 30 min. Further, these mixtures were used to cast 100 microns films on the metal/glass panels and subjected to cure at 110o C for 1 h and post-cure at 80 oC for 5 h. The nanocomposite surface coating film samples were tested for mechanical properties (viz. impact resistance, scratch hardness, adhesion, and flexibility) and resistance to artificial seawater (ASW). It was observed that 4 wt% nano-CaCO3 containing film sample showed maximum impact strength, scratch hardness, adhesion, and flexibility, and resistance to ASW compared to neat epoxy and other nanocomposite film samples. The curing behavior of nanocomposite film samples was analyzed by DSC. The reactivity effect of nano-CaCO3 blending on epoxy resin by FTIR spectroscopy, morphological studies by SEM, and surface gloss by Triglossometer were also investigated.

The low-density lipoprotein receptor (LDLR) controls cellular delivery of cholesterol and clears LDL from the bloodstream, protecting against atherosclerotic heart disease, the leading cause of death in the United States. We therefore sought to identify regulators of the LDLR beyond the targets of current therapies and known causes of familial hypercholesterolemia. We found that cold shock domain–containing protein E1 (CSDE1) enhanced hepatic LDLR messenger RNA (mRNA) decay via its 3′ untranslated region and regulated atherogenic lipoproteins in vivo. Using parallel phenotypic genome-wide CRISPR interference screens in a tissue culture model, we identified 40 specific regulators of the LDLR that were not previously identified by observational human genetic studies. Among these, we demonstrated that, in HepG2 cells, CSDE1 regulated the LDLR at least as strongly as statins and proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitors. In addition, we showed that hepatic gene silencing of Csde1 treated diet-induced dyslipidemia in mice to a similar degree as Pcsk9 silencing. These results suggest the therapeutic potential of targeting CSDE1 to manipulate the posttranscriptional regulation of the LDLR mRNA for the prevention of cardiovascular disease. Our approach of modeling a clinically relevant phenotype in a forward genetic screen, followed by mechanistic pharmacologic dissection and in vivo validation, may serve as a generalizable template for the identification of therapeutic targets in other human disease states.

Transcriptional networks governing cardiac precursor cell (CPC) specification are incompletely understood due in part to limitations in distinguishing CPCs from non-cardiac mesoderm in early gastrulation. We leveraged detection of early cardiac lineage transgenes within a granular single cell transcriptomic time course of mouse embryos to identify emerging CPCs and describe their transcriptional profiles. Mesp1, a transiently-expressed mesodermal transcription factor (TF), is canonically described as an early regulator of cardiac specification. However, we observed perdurance of CPC transgene-expressing cells in Mesp1 mutants, albeit mis-localized, prompting us to investigate the scope of Mesp1’s role in CPC emergence and differentiation. Mesp1 mutant CPCs failed to robustly activate markers of cardiomyocyte maturity and critical cardiac TFs, yet they exhibited transcriptional profiles resembling cardiac mesoderm progressing towards cardiomyocyte fates. Single cell chromatin accessibility analysis defined a Mesp1-dependent developmental breakpoint in cardiac lineage progression at a shift from mesendoderm transcriptional networks to those necessary for cardiac patterning and morphogenesis. These results reveal Mesp1-independent aspects of early CPC specification and underscore a Mesp1-dependent regulatory landscape required for progression through cardiogenesis.

Communication between myriad cell types during organ formation underlies proper morphogenesis1. In cardiac development, reciprocal signaling between mesoderm progenitors and neural crest cells is essential, and its disruption leads to congenital heart malformations, the most common human birth defect. However, mechanistic interrogation of temporal gene networks andcisregulatory elements in this crosstalk is limited2,3. Here, we integrated single cell chromatin accessibility and transcriptomics to establish an unbiased and temporal epigenomic map of the embryonic mouse heart over multiple stages and developed machine learning models to predict enhancers for heart and neural crest. We leveraged these advances to determine the consequences of dysregulated signaling at single cell resolution caused by deletion of TBX1, a transcription factor that causes morphogenetic defects of the cardiac outflow tract in humans and functions non-cell autonomously in cardiac mesodermal progenitors to direct pharyngeal neural crest differentiation4–6. Loss of Tbx1 in mice led to broad closure of chromatin regions enriched in cardiac progenitor transcription factor motifs within a narrow subset of cardiac mesodermal progenitors and correlated with diminished expression of numerous members of the fibroblast growth factor, retinoic acid, Notch and Semaphorin pathways. In affected progenitors, ectopic accessibility and expression of posterior heart field factors in the anterior heart field suggested impaired axial patterning. In response, a subset of cardiac neural crest cells displayed epigenomic and transcriptional defects, indicating a failure of differentiation corresponding to dysregulation of the anterior-posterior gradient of pharyngeal Hox gene expression. This study demonstrates that single-cell genomics and machine learning can generate a mechanistic model for how disruptions in cell communication selectively affect spatiotemporally dynamic regulatory networks in cardiogenesis.

Birth defects occur in ∼6% of all live births and can be caused by combinations of genetic and environmental influences1. Large-scale DNA sequencing efforts are revealing genetic influences2,3, but investigations into the contributions of environmental factors have largely been limited to association studies with limited mechanistic insight. Hyperglycemia present in pre-gestational diabetic mothers is among the most frequent environmental contributor to congenital defects and results in an increased incidence of congenital heart defects and craniofacial anomalies4. However, the cell types involved and underlying mechanisms by which maternal hyperglycemia affects these regions are unknown. Here, we utilized multi-modal single cell analyses to reveal that maternal diabetes affects the epigenomic and transcriptomic state of specific subsets of cardiac and craniofacial progenitors during embryogenesis. A previously unrecognized subpopulation of anterior heart field progenitors expressing Alx3 acquired a more posterior identity in response to maternal hyperglycemia, based on gene expression and chromatin status. Similarly, a sub-population of neural crest-derived cells in the second pharyngeal arch, which contributes to craniofacial structures, also displayed abnormalities in cell specification and patterning. Analysis of differentially accessible chromatin regions demonstrated that disrupted patterning was associated with increased intrinsic retinoic acid signaling in affected cell types in response to maternal diabetes and hyperglycemia. This work demonstrates how an environmental insult can have highly selective epigenomic consequences on discrete cell types leading to developmental patterning defects.

Drug repurposing processing plays a significant role in evaluating Drug Target identification in the field of Drug discovery. Today Machine learning algorithms based automated systems are gaining attention for creating new device applications in the field of artificial intelligence. [2] The present study used machine learning approaches to develop a computational predicted model for predicting the unknown pharmacological effects with immune-oncology compound. The authors applied similarity features on drug and disease to see indication also authors observed that if number of descriptor is increased then accuracy of model is also increased. Classification linear and nonlinear algorithms such as logistic regression,Support Vector Machine (Kernel) and Random Forest were used to build the computational models. We used 10 fold Cross-validation on dataset for train and test data to performed best (Accuracy = 0.932, AUC-ROC = 0.971), finally, the developed model was used to anticipate probable indications for currently available medications for cancer protein.

The well-established manifestation of mitochondrial mutations in functional cardiac disease (e.g., mitochondrial cardiomyopathy) prompted the hypothesis that mitochondrial DNA (mtDNA) sequence and/or copy number (mtDNAcn) variation contribute to cardiac defects in congenital heart disease (CHD). MtDNAcns were calculated and rare, non-synonymous mtDNA mutations were identified in 1,837 CHD-affected proband-parent trios, 116 CHD-affected singletons, and 114 paired cardiovascular tissue/blood samples. The variant allele fraction (VAF) of heteroplasmic variants in mitochondrial RNA from 257 CHD cardiovascular tissue samples was also calculated. On average, mtDNA from blood had 0.14 rare variants and 52.9 mtDNA copies per nuclear genome per proband. No variation with parental age at proband birth or CHD-affected proband age was seen. mtDNAcns in valve/vessel tissue (320 ± 70) were lower than in atrial tissue (1,080 ± 320, p= 6.8E-21), which were lower than in ventricle tissue (1,340 ± 280, p= 1.4E-4). The frequency of rare variants in CHD-affected individual DNA was indistinguishable from the frequency in an unaffected cohort, and proband mtDNAcns did not vary from those of CHD cohort parents. In both the CHD and the comparison cohorts, mtDNAcns were significantly correlated between mother-child, father-child, and mother-father. mtDNAcns among people with European (mean= 52.0), African (53.0), and Asian haplogroups (53.5) were calculated and were significantly different for European and Asian haplogroups (p= 2.6E-3). Variant heteroplasmic fraction (HF) in blood correlated well with paired cardiovascular tissue HF (r= 0.975) and RNA VAF (r= 0.953), which suggests blood HF is a reasonable proxy for HF in heart tissue. We conclude that mtDNA mutations and mtDNAcns are unlikely to contribute significantly to CHD risk.

Delimiting the Aravalli mountain range in the east, the Great Boundary Fault (GBF) occurs as a crustal-scale tectonic lineament in the NW Indian Shield. The structural and tectonic characteristics of the GBF are, as yet, not well-understood. We attempt to fill this gap by using a combination of satellite image processing, high-resolution outcrop mapping and structural analysis around Chittaurgarh. The study area exposes the core and damage zone of the GBF. Three successive phases of folding, F1, F2 and F3, are associated with deformation in the GBF. The large-scale structural characteristics of the GBF core are: (i) a non-coaxial refolding of F1 folds by F2 folds; and (ii) the parallelism between the GBF and F2 axial traces. In addition, numerous metre-scale ductile shear zones cut through the rocks in the GBF core. The damage zone is characterized by the large-scale F1 folds and the mesoscopic-scale strike-slip faults, thrusts and brittle-ductile shear zones. Several lines of evidence, such as the inconsistent overprinting relationship between the strike-slip faults and thrusts, the occurrence of en échelon folds and the palaeostress directions suggest that the GBF is a dextral transpression fault zone. Structural geometry and kinematic indicators imply a wrench- and contraction-dominated deformation in the core and damage zone, respectively. We infer that the GBF is a strain-partitioned dextral transpression zone.

Modeling Heart Development Engineering a model of the developing heart with stem cells has proved challenging. A recent study, however, provides a foundation — a single-chambered “cardioid” — for t...

Stokes drag on axially symmetric bodies vibrating slowly along the axis of symmetry placed under a uniform transverse flow of the Newtonian fluid is calculated. The axially symmetric bodies of revolution are considered with the condition of continuously turning tangent. The results of drag on sphere, spheroid, deformed sphere, egg-shaped body, cycloidal body, Cassini oval, and hypocycloidal body are found to be new. The numerical values of frictional drag on a slowly vibrating needle shaped body and flat circular disk are calculated as particular cases of deformed sphere.

Degenerative Scoliosis is well known for producing both neurological and mechanical symptoms in addition to producing deformity and sometimes instability. Decompression alone has been treatment of choice in patients with mild deformity with neurogenic claudication or radiculopathy but without instability. However, fusion has been preferred to counter deformity, instability and mechanical forces. We intended to study these variables when treated with decompression alone. We evaluated 38 patients who fulfilled our criteria. Demographic variables like age, gender, duration of symptoms and type of predominant symptom were noted. Clinical evaluation of back pain, deformity angle in supine and standing position was also performed both before and after surgery. Assessment was also done for type of symptoms and levels of decompression. We found no significant changes in deformity or back pain when decompression alone was done in such patients. The patients did improve symptomatically and neurologically. We found that decompression alone can be done to improve non-mechanical parameters in a patient of degenerative stable lumbar scoliosis.

Polymer nanocomposite samples were prepared by mechanical mixing of novolac epoxy polymer (EEW 180 g/eq) and varying concentrations of nano CaCO3 (0-5 wt%) for 30 min. using an electrical stirrer of 4000 rpm followed by uniform mixing of curing agent triethylenetetramine (TETA). The prepared mixtures were poured into a Teflon mold, designed as per ASTM standard. Thereafter subjected to cure at 110 °C for 1hr and post-cure at 70 °C for 5 hrs. The prepared nanocomposite samples were investigated for flexural strength, flexural modulus, impact strength, toughness, elongation at break, elastic modulus and hardness Shore D. It was observed that the sample containing 4 wt% nano CaCO3 showed the maximum value of flexural strength, flexural modulus, impact strength, toughness, elongation at break, elastic modulus and hardness Shore D, compared to the neat epoxy matrix. The curing reaction of nanocomposite samples was confirmed by FTIR spectroscopy and the morphological properties were investigated by SEM.

The modification of epoxy/fly ash composites by nano CaCO3 is an impressive way to enhance its prevailed properties. After modification of nano CaCO3, to understand mechanical behavior is meaningful in various fields, and enhancement in mechanical properties can advantageous to the convention in the various structural applications. The fly ash reinforced epoxy nanocomposite was prepared by mixing DGEBA epoxy resin and polyamine as a curing agent in which fly ash with 10 wt% and nano rCaCO3 with different (0, 1, 3 and 5) wt% were ultra-sonically dispersed for 30 min. The mechanical behavior of the prepared sample has been evaluated whereas the SEM technique was carried out to examine the morphological properties of epoxy/fly ash nanocomposite. Based on the morphological study, crack binding, crack transformation and strong interfacial bonding of CaCO3 nanoparticles were the prime reasons resulting in the enhancement of mechanical strength. The result indicates that the tensile strength of epoxy/fly ash composites increased by about 52.41% with the inclusion of 5 wt% CaCO3 nano reinforcement, whereas the impact strength and flexural strength were improved by 43.24% and 42.36% for the composition of 3 wt% nano CaCO3 reinforced epoxy/fly ash cast as compared to unmodified epoxy/fly ash cast. The thermal stability of epoxy/fly ash nanocomposites were analyzed by TGA and DSC technique. The degradation kinetics and water absorption behavior were also examined for above.

Highlights • Reduced AHN is linked closely with risk and resilience for psychiatric illness. • Prenatal MIA exposure reduces AHN through direct and indirect mechanisms. • We hypothesise that AHN may mediate susceptibility or resilience to MIA. • Studies in animal and human cellular models are required to test this hypothesis., Neurogenesis, the process in which new neurons are generated, occurs throughout life in the mammalian hippocampus. Decreased adult hippocampal neurogenesis (AHN) is a common feature across psychiatric disorders, including schizophrenia, depression- and anxiety-related behaviours, and is highly regulated by environmental influences. Epidemiological studies have consistently implicated maternal immune activation (MIA) during neurodevelopment as a risk factor for psychiatric disorders in adulthood. The extent to which the reduction of hippocampal neurogenesis in adulthood may be driven by early life exposures, such as MIA, is however unclear. We therefore reviewed the literature for evidence of the involvement of MIA in disrupting AHN. Consistent with our hypothesis, data from both in vivo murine and in vitro human models of AHN provide evidence for key roles of specific cytokines induced by MIA in the foetal brain in disrupting hippocampal neural progenitor cell proliferation and differentiation early in development. The precise molecular mechanisms however remain unclear. Nonetheless, these data suggest a potential latent vulnerability mechanism, whereby MIA primes dysfunction in the unique hippocampal pool of neural stem/progenitor cells. This renders offspring potentially more susceptible to additional environmental exposures later in life, such as chronic stress, resulting in the unmasking of psychopathology. We highlight the need for studies to test this hypothesis using validated animal models of MIA, but also to test the relevance of such data for human pathology at a molecular basis through the use of patient-derived induced pluripotent stem cells (hiPSC) differentiated into hippocampal progenitor cells.

This study reconstructs the pre-impact orientation of impact-generated µm-sized microstructures. The approach is illustrated using a sample from the basement dolerites of the Lockne impact structure, Sweden. Under the polarised light microscope, the microstructure appears as alternating augite-plagioclase wedges. The top-view of the microstructure is presented in the polished thin-section using light and scanning electron microscopy, while, the side-view is observed in focused-ionbeam trenches in a field emission scanning electron microscope. The top- and side view of the wedges are projected on a stereo net, which allows the determination of their 3D orientation. The wedges are thus found to have a dip of 08° towards 280°. The stereo net projections are then used to reconstruct the orientation of pre-impact boundary between augite-plagioclase, which had a dip of 15° towards 237°. This approach of combining light and electron microscopy with FIB trenching and projecting the data on a stereo net can be especially useful for reconstructing the pre-deformed state of impact-induced or tectonic µm-sized microstructures, which cannot be adequately analysed using a light microscope.

Exome sequencing of thousands of families has revealed many individual risk genes for congenital heart defects (CHD), yet most cases cannot be explained by a single causal mutation. Further, those who carryde novoand inherited mutations in known risk genes often demonstrate variable phenotypes even within the same family, indicating the presence of genetic modifiers. To explore oligogenic causes of CHD without assessing billions of variant combinations, we developed an efficient, simulation-based method to detect gene sets that carry damaging variants in probands at a higher rate than expected given parental genotypes. We implemented this approach in software called Gene Combinations in Oligogenic Disease (GCOD) and applied it to a cohort of 3382 trios with exome sequencing. This analysis detected 353 high-confidence risk genes in 202 pairs that appear together in multiple probands but rarely or never appear in combination in their unaffected parents. Stratifying analyses by specific CHD diagnosis and considering gene combinations of higher orders yielded an additional 244 gene sets. The oligogenic genes we discovered cluster in pathways specific to heart development and suggest new molecular disease mechanisms, such as arylsulfatase activity andde novonucleotide biosynthesis. Finally, by combining CHD families with an autism spectrum disorder cohort, we were able to detect 925 oligogenic sets transmitted in renal disease, a known co-morbidity of both conditions. As genome sequencing is applied to more families and other disorders, GCOD will enable detection of increasingly large, novel gene combinations, shedding light on combinatorial causes of genetic diseases.

Background: The regenerative capacity of the heart after myocardial infarction is limited. Our previous study showed that ectopic introduction of 4 cell cycle factors (4F; CDK1 [cyclin-dependent kinase 1], CDK4 [cyclin-dependent kinase 4], CCNB [cyclin B1], and CCND [cyclin D1]) promotes cardiomyocyte proliferation in 15% to 20% of infected cardiomyocytes in vitro and in vivo and improves cardiac function after myocardial infarction in mice. Methods: Using temporal single-cell RNA sequencing, we aimed to identify the necessary reprogramming stages during the forced cardiomyocyte proliferation with 4F on a single cell basis. Using rat and pig models of ischemic heart failure, we aimed to start the first preclinical testing to introduce 4F gene therapy as a candidate for the treatment of ischemia-induced heart failure. Results: Temporal bulk and single-cell RNA sequencing and further biochemical validations of mature human induced pluripotent stem cell–derived cardiomyocytes treated with either LacZ or 4F adenoviruses revealed full cell cycle reprogramming in 15% of the cardiomyocyte population at 48 hours after infection with 4F, which was associated mainly with sarcomere disassembly and metabolic reprogramming (n=3/time point/group). Transient overexpression of 4F, specifically in cardiomyocytes, was achieved using a polycistronic nonintegrating lentivirus (NIL) encoding 4F; each is driven by a TNNT2 (cardiac troponin T isoform 2) promoter (TNNT2-4Fpolycistronic-NIL). TNNT2-4Fpolycistronic-NIL or control virus was injected intramyocardially 1 week after myocardial infarction in rats (n=10/group) or pigs (n=6–7/group). Four weeks after injection, TNNT2-4Fpolycistronic-NIL–treated animals showed significant improvement in left ventricular ejection fraction and scar size compared with the control virus–treated animals. At 4 months after treatment, rats that received TNNT2-4Fpolycistronic-NIL still showed a sustained improvement in cardiac function and no obvious development of cardiac arrhythmias or systemic tumorigenesis (n=10/group). Conclusions: This study provides mechanistic insights into the process of forced cardiomyocyte proliferation and advances the clinical feasibility of this approach by minimizing the oncogenic potential of the cell cycle factors owing to the use of a novel transient and cardiomyocyte-specific viral construct.

Understanding how the geological architecture of the Himalaya has been constructed demands well-constrained tectonic models supported and validated by field observations. The channel flow model has been used to explain the structural architecture in many different sectors of the Himalaya. However, the model appeared to have failed in the classic example of the Himachal Himalaya, so in 2007 Webb proposed the Tectonic Wedge Model. But the key to this model is the existence of a regional-scale recumbent anticline, illustrated in many different papers and in textbooks. This structure is known variously as the Phojal fold, or the Sikhar nappe, or the Kalath anticline. Webb’s Tectonic Wedge Model requires the South Tibetan Detachment (STD) to also be recumbently folded, along with the Phojal fold. Our detailed field observations are contrary to the regional structures proposed in the existing models. First, it is evident that ‘photo geology’ has produced an optical illusion. Field mapping shows that the Phojal Fold is a north-east vergent reclined back fold. Thus, despite having been developed on the km-scale, the Phojal Fold has nothing to do with the formation of the earlier formed recumbent folds. There is no doubt that an early period of recumbent folding has produced regional-scale structures in the NW Himalaya. These folds post-date the first recognized period of Barrovian metamorphism. However, because the axial-plane cleavage of the recumbent folds is a pressure-solution cleavage, it can be inferred that these metamorphic rocks had cooled and been exhumed to shallow crustal levels prior to the start of the early recumbent folding event. The second period of Barrovian metamorphism was associated with the STD, which post-dates the earlier recumbent folds, but pre-dates back folding. The underpinning of the Tectonic Wedge Model has been removed. Hence the validity of the model is on trial.

SummaryFemale mammalian cells harbor two X chromosomes, one of maternal and one of paternal origin. During development, one X randomly inactivates1–4. This renders either the maternal or paternal X active, causing X mosaicism that varies among individual females, with some showing considerable or complete skew in the general female population5–7. Parent-of-X-origin can modify epigenetics via DNA methylation8, 9 and possibly gene expression; thus, mosaicism could buffer dysregulated processes in aging and disease. However, whether X skewing – or its mosaicism – alters functions in females is largely unknown. Here we tested whether skew toward the maternal X (Xm) influences key functions of the body. Among cardiac, bone, metabolic, and brain functions, Xm selectively impaired cognition in female mice throughout the lifespan. Cognitive deficits were accompanied by Xm-mediated acceleration of biologic or epigenetic aging of the female hippocampus, a key center for learning and memory. Xm showed epigenetic imprinting of several genes within hippocampal neurons, suggesting silenced cognitive loci. Thus, the maternal X chromosome impaired cognition, accelerated brain aging, and silenced genes. Understanding how the maternal X impairs brain function could lead to new understanding of female heterogeneity in cognitive heath and to X chromosome-derived pathways against cognitive deficits and brain aging.

Congenital heart disease (CHD) is present in 1% of live births, yet identification of causal mutations remains challenging. We hypothesized that genetic determinants for CHDs may lie in the protein interactomes of transcription factors whose mutations cause CHDs. Defining the interactomes of two transcription factors haplo-insufficient in CHD, GATA4 and TBX5, within human cardiac progenitors, and integrating the results with nearly 9,000 exomes from proband-parent trios revealed an enrichment of de novo missense variants associated with CHD within the interactomes. Scoring variants of interactome members based on residue, gene, and proband features identified likely CHD-causing genes, including the epigenetic reader GLYR1. GLYR1 and GATA4 widely co-occupied and co-activated cardiac developmental genes, and the identified GLYR1 missense variant disrupted interaction with GATA4, impairing invitro and invivo function in mice. This integrative proteomic and genetic approach provides a framework for prioritizing and interrogating genetic variants in heart disease.

Increased thoracic kyphosis (hyperkyphosis) is well known entity especially in elderly population may be responsible for multiple morbidity and needs a non-pharmacological preventive strategy due to which muscle strength training has been routinely advised. Eighty patients were included in study and divided into two groups. One group received institutional and home-based exercise regimen and another group was administered the routine pharmacological regimen. Besides demographic variables, clinical (occiput to wall distance, pain, ODI score and spinal flexibility) and radiological parameters (Cobb’s angles of thoracic kyphosis, lumbar lordosis, and thoracic kyphosis in best erect posture) were assessed before and after the end of 3-month duration. The data between both the groups was comparable but the study group showed marked improvement at the end of study (mean difference of 1.8o) and significant improvement in clinical parameters of pain, ODI score, occiput to wall distance and spinal flexibility. The other values were insignificantly affected. We find that exercise regimen positively impacts and significantly improves the thoracic kyphosis, clinical and functional status of patients who have hyperkyphosis but undergo a targeted muscle training programme for back muscles.

Today’s ‘gurus’ have stolen the show in the spiritual and commercial arena by making yoga and Ayurveda popular. This case study discusses the strategic marketing mix strategy adopted by Patanjali Ayurved Limited and Sri Sri Tattva. The case study brings forward the statistical underpinnings of both companies and carefully presents the qualitative assessment of the moves and countermoves of these companies against their respective set of competitors in the market. Moreover, the case study raises some pertinent questions that these companies must consider remaining in the market and retain their competitive edge.

A genetic etiology is identified for one-third of patients with congenital heart disease (CHD), with 8% of cases attributable to coding de novo variants (DNVs). To assess the contribution of noncoding DNVs to CHD, we compared genome sequences from 749 CHD probands and their parents with those from 1,611 unaffected trios. Neural network prediction of noncoding DNV transcriptional impact identified a burden of DNVs in individuals with CHD (n = 2,238 DNVs) compared to controls (n = 4,177; P = 8.7 × 10-4). Independent analyses of enhancers showed an excess of DNVs in associated genes (27 genes versus 3.7 expected, P = 1 × 10-5). We observed significant overlap between these transcription-based approaches (odds ratio (OR) = 2.5, 95% confidence interval (CI) 1.1-5.0, P = 5.4 × 10-3). CHD DNVs altered transcription levels in 5 of 31 enhancers assayed. Finally, we observed a DNV burden in RNA-binding-protein regulatory sites (OR = 1.13, 95% CI 1.1-1.2, P = 8.8 × 10-5). Our findings demonstrate an enrichment of potentially disruptive regulatory noncoding DNVs in a fraction of CHD at least as high as that observed for damaging coding DNVs.

COVID-19 shutdowns have impacted research institutions across the globe. Cell Stem Cell asked Deepak Srivastava, Fiona Watt, and George Daley to share their perspectives on leadership during this crisis and how their institutions are supporting the scientific community to address this global health crisis.