Thulaj Dattatraya Meharwade

All Publications

• Cell Village Captures Populational Heterogeneity of Sunitinib Cardiotoxicity. Circulation research Kojic, A., Liu, Y., Belbachir, N., Tu, C., Shen, M., Caudal, A., Meharwade, T., Belbachir, M., Ford, K., Sallam, K., Wu, J. C. 2026

  Abstract

  TKIs (tyrosine kinase inhibitors) have transformed cancer therapy; however, cardiotoxicity remains a frequent adverse effect with variable susceptibility among patients. For sunitinib, a commonly prescribed TKI, the molecular programs and functional features underlying this variability are not well defined, and clinically actionable predictors of cardiotoxicity risk remain lacking.To investigate interindividual variability in sunitinib-induced cardiotoxic responses, we implemented the Cell Village platform consisting of human induced pluripotent stem cell (iPSC)-derived cardiomyocytes from 20 healthy donors. Pooled iPSCs were differentiated into cardiomyocytes, treated with sunitinib for 72 hours, followed by harvesting for single-nucleus RNA sequencing. Donor identity was determined using single nucleotide polymorphism-based demultiplexing. Additionally, individual line-specific transcriptomic responses to sunitinib were integrated with quantitative functional phenotyping (including contractility, electrophysiology, and recovery efficacy following tocotrienol treatment) in iPSC-derived cardiomyocytes differentiated from a subset of 10 lines.Sunitinib induced a robust global transcriptional response characterized by dysregulation of tyrosine kinase signaling, cytoskeletal organization, ion channels, and calcium-handling pathways. Although the pathway-level changes were shared, individual donor lines displayed heterogeneity in both gene expression patterns and functional impairment. Across functional parameters, relaxation velocity was the most consistently correlated with toxicity severity. Integration of each iPSC line's functional data with the corresponding transcriptomic signatures identified gene modules related to calcium handling and structural organization strongly correlated with donor-specific toxicity and recovery indices. Pharmacological validation using tocotrienol demonstrated partial, donor-dependent improvement of sunitinib-related iPSC-derived cardiomyocyte impairments.This proof-of-concept study establishes a scalable, donor-resolved platform that links pooled transcriptomic profiling with functional toxicity and recovery phenotypes in human cardiomyocytes. By linking molecular perturbations to functional outcomes across genetically diverse backgrounds, this approach provides a framework for dissecting interindividual variability in TKI-associated cardiotoxicity. More broadly, it provides a foundation for extending population-scale cardiotoxicity assessment to additional drugs and protective interventions.

  View details for DOI 10.1161/CIRCRESAHA.126.328704

  View details for PubMedID 42852499

• Generation of two induced pluripotent stem cell lines from female patients harboring MYH7 mutations with hypertrophic cardiomyopathy. Stem cell research Meharwade, T., Weiss, M. C., Ahmed, A. H., Xu, L., Parikh, V. N., Ashley, E. A., Wu, J. C. 2026; 94: 103979

  Abstract

  Hypertrophic cardiomyopathy (HCM) is a cardiac disorder marked by left ventricular thickening and impaired diastolic function, often caused by truncating variants in MYH7 gene. In this work, we generated two human iPSC lines from female HCM patients, each carrying a distinct heterozygous MYH7 mutation (c.611G > T and c.505A > G). Both lines displayed a normal karyotype, typical pluripotent stem cell morphology, strong expression of key pluripotency markers, and retained the capacity to differentiate into cell types representing all three germ layers. These patient-specific female iPSC lines provide a valuable platform to explore inter-patient variability and the mechanistic diversity underlying MYH7-associated cardiomyopathies.

  View details for DOI 10.1016/j.scr.2026.103979

  View details for PubMedID 41985226

• Cross-activation of FGF, NODAL, and WNT pathways constrains BMP-signaling-mediated induction of the totipotent state in mouse embryonic stem cells CELL REPORTS Meharwade, T., Joumier, L., Parisotto, M., Huynh, V., da Rocha, E., Malleshaiah, M. 2023; 42 (5): 112438

  Abstract

  Embryonic stem cells (ESCs) are an attractive model to study the relationship between signaling and cell fates. Cultured mouse ESCs can exist in multiple states resembling distinct stages of early embryogenesis, such as totipotent, pluripotent, primed, and primitive endoderm. The signaling mechanisms regulating the totipotent state and coexistence of these states are poorly understood. Here we identify bone morphogenetic protein (BMP) signaling as an inducer of the totipotent state. However, we discover that BMP's role is constrained by the cross-activation of FGF, NODAL, and WNT pathways. We exploit this finding to enhance the proportion of totipotent cells by rationally inhibiting the cross-activated pathways. Single-cell mRNA sequencing reveals that induction of the totipotent state is accompanied by suppression of primed and primitive endoderm states. Furthermore, reprogrammed totipotent cells we generate in culture resemble totipotent cells of preimplantation embryo. Our findings reveal a BMP signaling mechanism regulating both the totipotent state and heterogeneity of ESCs.

  View details for DOI 10.1016/j.celrep.2023.112438

  View details for Web of Science ID 000990371100001

  View details for PubMedID 37126449

• Single-cell mass cytometry analysis reveals stem cell heterogeneity. Methods (San Diego, Calif.) Meharwade, T., Joumier, L., Parisotto, M., Malleshaiah, M. 2022; 208: 9-18

  Abstract

  Cellular heterogeneity is fundamental to both developmental differentiation and disease establishment. Recent advances in high-throughput single-cell technology have been rapidly revolutionizing the resolution of our understanding of development and disease. However, while the study of single-cell transcriptomes is easily accessible, the analysis of single-cell proteomes is still in its infancy. In this study, we describe simultaneous profiling of multiple regulatory proteins at a single-cell level using mass cytometry or cytometry by time of flight. We develop mass cytometry reagents to study key transcription factors, signaling proteins and chromatin modifiers that regulate mouse embryonic stem cells. Our data reveal that the protein level of stem cell regulators significantly varies and that cell signaling pathways are extensively cross-activated across defined culture conditions of embryonic stem cells. In addition, the mass cytometry data enabled us to identify distinct multiple cell states of embryonic stem cells and determine their variation across culture conditions. We discuss the mass cytometry method, our results of the multi-protein analysis in embryonic stem cells and potential future perspectives for single-cell protein analysis.

  View details for DOI 10.1016/j.ymeth.2022.09.009

  View details for PubMedID 36229002

• The NAMPT Inhibitor FK866 Increases Metformin Sensitivity in Pancreatic Cancer Cells CANCERS Parisotto, M., Vuong-Robillard, N., Kalegari, P., Meharwade, T., Joumier, L., Igelmann, S., Bourdeau, V., Rowell, M., Pollak, M., Malleshaiah, M., Schmitzer, A., Ferbeyre, G. 2022; 14 (22)

  Abstract

  Pancreatic cancer (pancreatic ductal adenocarcinoma: PDAC) is one of the most aggressive neoplastic diseases. Metformin use has been associated with reduced pancreatic cancer incidence and better survival in diabetics. Metformin has been shown to inhibit PDAC cells growth and survival, both in vitro and in vivo. However, clinical trials using metformin have failed to reduce pancreatic cancer progression in patients, raising important questions about molecular mechanisms that protect tumor cells from the antineoplastic activities of metformin. We confirmed that metformin acts through inhibition of mitochondrial complex I, decreasing the NAD+/NADH ratio, and that NAD+/NADH homeostasis determines metformin sensitivity in several cancer cell lines. Metabolites that can restore the NAD+/NADH ratio caused PDAC cells to be resistant to metformin. In addition, metformin treatment of PDAC cell lines induced a compensatory NAMPT expression, increasing the pool of cellular NAD+. The NAMPT inhibitor FK866 sensitized PDAC cells to the antiproliferative effects of metformin in vitro and decreased the cellular NAD+ pool. Intriguingly, FK866 combined with metformin increased survival in mice bearing KP4 cell line xenografts, but not in mice with PANC-1 cell line xenografts. Transcriptome analysis revealed that the drug combination reactivated genes in the p53 pathway and oxidative stress, providing new insights about the mechanisms leading to cancer cell death.

  View details for DOI 10.3390/cancers14225597

  View details for Web of Science ID 000887064200001

  View details for PubMedID 36428689

  View details for PubMedCentralID PMC9688551