Nadjet Belbachir

All Publications

• Empagliflozin Attenuates Arrhythmias in an iPSC-Based Model of Hypertrophy Cardiomyopathy. Circulation. Genomic and precision medicine Arthur Ataam, J., Belbachir, N., Perea-Gil, I., Termglincan, V., Vadgama, N., Garg, P., Ramchandani, R., Gavidia, A. A., Roura, S., Gálvez-Montón, C., Wu, J. C., Bayés-Genis, A., Karakikes, I. 2024: e004526

  View details for DOI 10.1161/CIRCGEN.123.004526

  View details for PubMedID 38752368

• Generation of induced pluripotent stem cell lines from patients with LQT1 caused by heterozygous mutations in the KCNQ1 gene. Stem cell research Ren, L., Jahng, J. W., Belbachir, N., Cook, Z., Rivero, G. C., Perez, M. V., Wu, J. C. 2024; 78: 103443

  Abstract

  Long QT Syndrome (LQTS) is a genetic heart disorder that can induce cardiac arrhythmias. The most prevalent subtype, LQT1, stems from rare variants in the KCNQ1 gene. Utilizing induced pluripotent stem cells (iPSCs) enables detailed cellular studies and personalized medicine approaches for this life-threatening condition. We generated two LQT1 iPSC lines with single nucleotide nonsense mutations, c.1031 C>T and c.1121T>A in KCNQ1. Both lines exhibited typical iPSC morphology, expressed high levels of pluripotent markers, maintained normal karyotype, and possessed the capability to differentiate into three germ layers. These cell lines serve as important tools for investigating the biological mechanisms underlying LQT1 due to mutations in the KCNQ1 gene.

  View details for DOI 10.1016/j.scr.2024.103443

  View details for PubMedID 38763038

• Studying Long QT Syndrome Caused by NAA10 Genetic Variants Using Patient-Derived Induced Pluripotent Stem Cells. Circulation Belbachir, N., Wu, Y., Shen, M., Zhang, S. L., Zhang, J. Z., Liu, C., Knollmann, B. C., Lyon, G. J., Ma, N., Wu, J. C. 2023; 148 (20): 1598-1601

  View details for DOI 10.1161/CIRCULATIONAHA.122.061864

  View details for PubMedID 37956223

• Generation of two induced pluripotent stem cell lines from catecholaminergic polymorphic ventricular tachycardia patients carrying RYR2 mutations. Stem cell research Kong, X., Belbachir, N., Zeng, W., Yan, C. D., Navada, S., Perez, M. V., Wu, J. C. 2023; 69: 103111

  Abstract

  Catecholaminergic polymorphic ventricular tachycardia (CPVT) is a congenital arrhythmic syndrome caused by the RYR2 gene encoded ryanodine receptor. Mutations on RYR2 are commonly associated with ventricular tachycardia after adrenergic stimulation, leading to lethal arrhythmias and sudden cardiac death. We generated two human induced pluripotent stem cell (iPSC) lines from CPVT affected patients carrying single missense heterozygote RYR2 mutations, c.1082 G > A and c.100 A > C. Pluripotency and differentiation capability into derivatives of three germ layers were evaluated along with karyotype stability in the report. The generated patient-specific iPSC lines provide a reliable tool to investigate the CPVT phenotype and understand underlaying mechanisms.

  View details for DOI 10.1016/j.scr.2023.103111

  View details for PubMedID 37210947

• High-Throughput Analysis of Drug Safety Responses in Induced Pluripotent Stem Cell-Derived Cardiomyocytes Using Multielectrode Array. Methods in molecular biology (Clifton, N.J.) Belbachir, N., Cunningham, N., Wu, J. C. 2022; 2485: 99-109

  Abstract

  Microelectrode array (MEA) is an electrophysiological instrument used to track activities of ion channels in excitable cells. Neurons and cardiomyocytes are seeded to form a cell monolayer on a field of sensors able to detect electrical signals, called extracellular field potentials (EFPs). This noninvasive tool allows researchers to investigate key parameters such as EFP amplitude, duration, and arrhythmias. MEA is progressively considered the gold standard for high-throughput in vitro electrophysiological evaluation, particularly for cardiac disease modeling and cardiac toxicity assessment.

  View details for DOI 10.1007/978-1-0716-2261-2_7

  View details for PubMedID 35618901

• Generation of human induced pluripotent stem cell lines from four unrelated healthy control donors carrying European genetic background. Stem cell research Girardeau, A., Atticus, D., Canac, R., Cimarosti, B., Caillaud, A., Chariau, C., Simonet, F., Cariou, B., Charpentier, F., Gourraud, J., Probst, V., Belbachir, N., Jesel, L., Lemarchand, P., Barc, J., Redon, R., Gaborit, N., Lamirault, G. 1800; 59: 102647

  Abstract

  Four human induced pluripotent stem cell (hiPSC) lines have been generated from healthy control European donors, and validated. This resource represents a useful tool for stem cell-based research, as references for developmental studies and disease modeling linked to any type of human tissue and organ, in an ethnical-, sex- and age-matched context. They providea reliable in-vitro model for single cell- and tissue-based investigations, and are also a valuable tool for genome editing-based studies.

  View details for DOI 10.1016/j.scr.2021.102647

  View details for PubMedID 34999420

• Generation of two induced pluripotent stem cell lines from Brugada syndrome affected patients carrying SCN5A mutations. Stem cell research Belbachir, N., Lai, C., Rhee, J., Zhuge, Y., Perez, M. V., Sallam, K., Wu, J. C. 2021; 57: 102605

  Abstract

  SCN5A gene loss-of-function mutations are commonly associated with Brugada syndrome, which represents a risk of lethal arrhythmias and sudden cardiac death. The present report describes the generation of two human induced pluripotent stem cell (iPSC) lines reprogrammed from two Brugada syndrome affected patients carrying SCN5A mutations, c.53506 G>A and c.2102 C>T, respectively. Pluripotency markers, karyotype stability, and differentiation capability into derivatives of the three germ layers were assessed and described in the present report. These lines can be used as a reliable cell model for Brugada syndrome investigations and characterization of leading cellular mechanisms.

  View details for DOI 10.1016/j.scr.2021.102605

  View details for PubMedID 34856468

• Endocardial/endothelial angiocrines regulate cardiomyocyte development and maturation and induce features of ventricular non-compaction. European heart journal Rhee, S., Paik, D. T., Yang, J. Y., Nagelberg, D., Williams, I., Tian, L., Roth, R., Chandy, M., Ban, J., Belbachir, N., Kim, S., Zhang, H., Phansalkar, R., Wong, K. M., King, D. A., Valdez, C., Winn, V. D., Morrison, A. J., Wu, J. C., Red-Horse, K. 2021

  Abstract

  AIMS: Non-compaction cardiomyopathy is a devastating genetic disease caused by insufficient consolidation of ventricular wall muscle that can result in inadequate cardiac performance. Despite being the third most common cardiomyopathy, the mechanisms underlying the disease, including the cell types involved, are poorly understood. We have previously shown that endothelial cell-specific deletion of the chromatin remodeller gene Ino80 results in defective coronary vessel development that leads to ventricular non-compaction in embryonic mouse hearts. We aimed to identify candidate angiocrines expressed by endocardial and ECs inwildtype and LVNC conditions in Tie2Cre;Ino80fl/fl transgenic embryonic mouse hearts, and test the effect of these candidates on cardiomyocyte proliferation and maturation.METHODS AND RESULTS: We used single-cell RNA-sequencing to characterize endothelial and endocardial defects in Ino80-deficient hearts. We observed a pathological endocardial cell population in the non-compacted hearts and identified multiple dysregulated angiocrine factors that dramatically affected cardiomyocyte behaviour. We identified Col15A1 as a coronary vessel-secreted angiocrine factor, downregulated by Ino80-deficiency, that functioned to promote cardiomyocyte proliferation. Furthermore, mutant endocardial and endothelial cells (ECs) up-regulated expression of secreted factors, such as Tgfbi, Igfbp3, Isg15, and Adm, which decreased cardiomyocyte proliferation and increased maturation.CONCLUSIONS: These findings support a model where coronary ECs normally promote myocardial compaction through secreted factors, but that endocardial and ECs can secrete factors that contribute to non-compaction under pathological conditions.

  View details for DOI 10.1093/eurheartj/ehab298

  View details for PubMedID 34279605

• Modeling Secondary Iron Overload Cardiomyopathy with Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes. Cell reports Rhee, J. W., Yi, H. n., Thomas, D. n., Lam, C. K., Belbachir, N. n., Tian, L. n., Qin, X. n., Malisa, J. n., Lau, E. n., Paik, D. T., Kim, Y. n., Choi, B. S., Sayed, N. n., Sallam, K. n., Liao, R. n., Wu, J. C. 2020; 32 (2): 107886

  Abstract

  Excessive iron accumulation in the heart causes iron overload cardiomyopathy (IOC), which initially presents as diastolic dysfunction and arrhythmia but progresses to systolic dysfunction and end-stage heart failure when left untreated. However, the mechanisms of iron-related cardiac injury and how iron accumulates in human cardiomyocytes are not well understood. Herein, using human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs), we model IOC and screen for drugs to rescue the iron overload phenotypes. Human iPSC-CMs under excess iron exposure recapitulate early-stage IOC, including oxidative stress, arrhythmia, and contractile dysfunction. We find that iron-induced changes in calcium kinetics play a critical role in dysregulation of CM functions. We identify that ebselen, a selective divalent metal transporter 1 (DMT1) inhibitor and antioxidant, could prevent the observed iron overload phenotypes, supporting the role of DMT1 in iron uptake into the human myocardium. These results suggest that ebselen may be a potential preventive and therapeutic agent for treating patients with secondary iron overload.

  View details for DOI 10.1016/j.celrep.2020.107886

  View details for PubMedID 32668256

• Levitating Cells to Sort the Fit and the Fat. Advanced biosystems Puluca, N. n., Durmus, N. G., Lee, S. n., Belbachir, N. n., Galdos, F. X., Ogut, M. G., Gupta, R. n., Hirano, K. I., Krane, M. n., Lange, R. n., Wu, J. C., Wu, S. M., Demirci, U. n. 2020: e1900300

  Abstract

  Density is a core material property and varies between different cell types, mainly based on differences in their lipid content. Sorting based on density enables various biomedical applications such as multi-omics in precision medicine and regenerative repair in medicine. However, a significant challenge is sorting cells of the same type based on density differences. Here, a new method for real-time monitoring and sorting of single cells based on their inherent levitation profiles driven by their lipid content is reported. As a model system, human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (CMs) from a patient with neutral lipid storage disease (NLSD) due to loss of function of adipose triglyceride lipase (ATGL) resulting in abnormal lipid storage in cardiac muscle are used. This levitation-based strategy detects subpopulations within ATGL-deficient hiPSC-CMs with heterogenous lipid content, equilibrating at different levitation heights due to small density differences. In addition, sorting of these differentially levitating subpopulations are monitored in real time. Using this approach, sorted healthy and diseased hiPSC-CMs maintain viability and function. Pixel-tracking technologies show differences in contraction between NLSD and healthy hiPSC-CMs. Overall, this is a unique approach to separate diseased cell populations based on their intracellular lipid content that cannot be achieved using traditional flow cytometry techniques.

  View details for DOI 10.1002/adbi.201900300

  View details for PubMedID 32352239

• Effects of Cryopreservation on Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes for Assessing Drug Safety Response Profiles. Stem cell reports Zhang, J. Z., Belbachir, N. n., Zhang, T. n., Liu, Y. n., Shrestha, R. n., Wu, J. C. 2020

  Abstract

  Burgeoning applications of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) in disease modeling, regenerative medicine, and drug screening have broadened the usage of hiPSC-CMs and entailed their long-term storage. Cryopreservation is the most common approach to store hiPSC-CMs. However, the effects of cryopreservation and recovery on hiPSC-CMs remain poorly understood. Here, we characterized the transcriptome, electro-mechanical function, and drug response of fresh hiPSC-CMs without cryopreservation and recovered hiPSC-CMs from cryopreservation. We found that recovered hiPSC-CMs showed upregulation of cell cycle genes, similar or reduced contractility, Ca2+ transients, and field potential duration. When subjected to treatment of drugs that affect electrophysiological properties, recovered hiPSC-CMs showed an altered drug response and enhanced propensity for drug-induced cardiac arrhythmic events. In conclusion, fresh and recovered hiPSC-CMs do not always show comparable molecular and physiological properties. When cryopreserved hiPSC-CMs are used for assessing drug-induced cardiac liabilities, the altered drug sensitivity needs to be considered.

  View details for DOI 10.1016/j.stemcr.2020.11.010

  View details for PubMedID 33338435

• Identifying the Transcriptome Signature of Calcium Channel Blocker in Human iPSC-Derived Cardiomyocytes Lam, C., Tian Lei, Belbachir, N., Wnorowski, A., Shrestha, R., Ma Ning, Kitani, T., Rhee, J. W., Wu, J. C. LIPPINCOTT WILLIAMS & WILKINS. 2019

  View details for Web of Science ID 000511467800108

• Identifying the Transcriptome Signatures of Calcium Channel Blockers in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes CIRCULATION RESEARCH Lam, C., Tian, L., Belbachir, N., Wnorowski, A., Shrestha, R., Ma, N., Kitani, T., Rhee, J., Wu, J. C. 2019; 125 (2): 212–22

  View details for DOI 10.1161/CIRCRESAHA.118.314202

  View details for Web of Science ID 000474146200010

• RRAD mutation causes electrical and cytoskeletal defects in cardiomyocytes derived from a familial case of Brugada syndrome. European heart journal Belbachir, N., Portero, V., Al Sayed, Z. R., Gourraud, J., Dilasser, F., Jesel, L., Guo, H., Wu, H., Gaborit, N., Guilluy, C., Girardeau, A., Bonnaud, S., Simonet, F., Karakachoff, M., Pattier, S., Scott, C., Burel, S., Marionneau, C., Chariau, C., Gaignerie, A., David, L., Genin, E., Deleuze, J., Dina, C., Sauzeau, V., Loirand, G., Baro, I., Schott, J., Probst, V., Wu, J. C., Redon, R., Charpentier, F., Le Scouarnec, S. 2019

  Abstract

  AIMS: The Brugada syndrome (BrS) is an inherited cardiac disorder predisposing to ventricular arrhythmias. Despite considerable efforts, its genetic basis and cellular mechanisms remain largely unknown. The objective of this study was to identify a new susceptibility gene for BrS through familial investigation.METHODS AND RESULTS: Whole-exome sequencing performed in a three-generation pedigree with five affected members allowed the identification of one rare non-synonymous substitution (p.R211H) in RRAD, the gene encoding the RAD GTPase, carried by all affected members of the family. Three additional rare missense variants were found in 3/186 unrelated index cases. We detected higher levels of RRAD transcripts in subepicardium than in subendocardium in human heart, and in the right ventricle outflow tract compared to the other cardiac compartments in mice. The p.R211H variant was then subjected to electrophysiological and structural investigations in human cardiomyocytes derived from induced pluripotent stem cells (iPSC-CMs). Cardiomyocytes derived from induced pluripotent stem cells from two affected family members exhibited reduced action potential upstroke velocity, prolonged action potentials and increased incidence of early afterdepolarizations, with decreased Na+ peak current amplitude and increased Na+ persistent current amplitude, as well as abnormal distribution of actin and less focal adhesions, compared with intra-familial control iPSC-CMs Insertion of p.R211H-RRAD variant in control iPSCs by genome editing confirmed these results. In addition, iPSC-CMs from affected patients exhibited a decreased L-type Ca2+ current amplitude.CONCLUSION: This study identified a potential new BrS-susceptibility gene, RRAD. Cardiomyocytes derived from induced pluripotent stem cells expressing RRAD variant recapitulated single-cell electrophysiological features of BrS, including altered Na+ current, as well as cytoskeleton disturbances.

  View details for DOI 10.1093/eurheartj/ehz308

  View details for PubMedID 31114854

• Identifying the Transcriptome Signatures of Calcium Channel Blockers in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes. Circulation research Lam, C. K., Tian, L. n., Belbachir, N. n., Shrestha, R. n., Ma, N. n., Kitani, T. n., Rhee, J. W., Wnorowski, A. n., Wu, J. C. 2019

  Abstract

  Calcium channel blockers (CCBs) are an important class of drugs in managing cardiovascular diseases. Patients usually rely on these medications for the remainder of their lives after diagnosis. Although the acute pharmacological actions of CCBs in the hearts are well-defined, little is known about the drug-specific effects on human cardiomyocyte transcriptomes and physiological alterations after long-term exposure.This study aimed to simulate chronic CCB treatment and to examine both the functional and transcriptomic changes in human cardiomyocytes.We differentiated cardiomyocytes and generated engineered heart tissues from three human induced pluripotent stem cell (iPSC) lines and exposed them to four different CCBs-nifedipine, amlodipine, diltiazem, and verapamil-at their physiological serum concentrations for two weeks. Without inducing cell death and damage to myofilament structure, CCBs elicited line-specific inhibition on calcium kinetics and contractility. While all four CCBs exerted similar inhibition on calcium kinetics, verapamil applied the strongest inhibition on cardiomyocyte contractile function. By profiling cardiomyocyte transcriptome after CCB treatment, we identified little overlap in their transcriptome signatures. Verapamil is the only inhibitor that reduced the expression of contraction-related genes, such as myosin heavy chain and troponin I, consistent with its depressive effects on contractile function. The reduction of these contraction related genes may also explain the responsiveness of HCM patients to verapamil in managing left ventricular outflow tract obstruction.This is the first study to identify the transcriptome signatures of different CCBs in human cardiomyocytes. The distinct gene expression patterns suggest that although the four inhibitors act on the same target, they may have distinct effects on normal cardiac cell physiology.

  View details for PubMedID 31079550