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Article ; Online: Human Induced Pluripotent Stem Cell-Derived Lung Epithelial System for SARS-CoV-2 Infection Modeling and Its Potential in Drug Repurposing.

Surendran, Harshini / Nandakumar, Swapna / Pal, Rajarshi

2020 Volume 29, Issue 21, Page(s) 1365–1369

Abstract: The lung is the most vulnerable target for the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection, and respiratory failure causing acute respiratory distress syndrome is its foremost outcome. However, the current primary in vitro ... ...

Abstract	The lung is the most vulnerable target for the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection, and respiratory failure causing acute respiratory distress syndrome is its foremost outcome. However, the current primary in vitro models in use for SARS-CoV-2 display apparent limitations for modeling such complex human respiratory disease. Although patient cells can directly model the effects of a drug, their availability and capacity for expansion are limited compared with transformed/immortalized cells or tumor-derived cell lines. An additional caveat is that the latter may harbor genetic and metabolic abnormalities making them unsuitable for drug screening. Therefore, it is important to create physiologically relevant human-cell models that can replicate the pathophysiology of SARS-CoV-2, thus facilitating drug testing. In this study, we show preliminary data on how human induced pluripotent stem cells-derived lung epithelial cell system could emerge as a relevant and sensitive platform for modeling SARS-CoV-2 infection and drug screening.
MeSH term(s)	COVID-19/drug therapy ; COVID-19/pathology ; Cell Line ; Drug Evaluation, Preclinical/methods ; Drug Repositioning ; Humans ; Induced Pluripotent Stem Cells/virology ; Lung/cytology ; Models, Biological ; Respiratory Distress Syndrome/pathology ; Respiratory Distress Syndrome/prevention & control ; SARS-CoV-2/drug effects
Keywords	covid19
Language	English
Publishing date	2020-09-18
Publishing country	United States
Document type	Letter ; Research Support, Non-U.S. Gov't
ZDB-ID	2142214-X
ISSN	1557-8534 ; 1547-3287
ISSN (online)	1557-8534
ISSN	1547-3287
DOI	10.1089/scd.2020.0152
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Article ; Online: Derivation of Induced Pluripotent Stem Cell (iPSC) Lines from Patient-Specific Peripheral Blood Mononuclear Cells (PBMC) Using Episomal Vectors.

Konala, Vijay Bhaskar Reddy / Nandakumar, Swapna / Surendran, Harshini / Pal, Rajarshi

Methods in molecular biology (Clifton, N.J.)

2021 Volume 2549, Page(s) 137–151

Abstract: Inherited retinal diseases (IRDs) are a diverse group of rare eye disorders, resulting in vision loss or blindness. The underlying reason is mutation in one or more than 250 different genes associated with the development and normal physiology of retina ... ...

Abstract	Inherited retinal diseases (IRDs) are a diverse group of rare eye disorders, resulting in vision loss or blindness. The underlying reason is mutation in one or more than 250 different genes associated with the development and normal physiology of retina largely comprising of rod/cone photoreceptors and retinal pigment epithelium. Interestingly, the sub retinal region of an eye has been shown to be immune privileged, broadening the scope of cell-replacement therapies for patients suffering from retinal degeneration. Several groups around the globe, including ours, have demonstrated safety and efficacy in preclinical studies by employing various approaches of retinal cell therapy. This had largely been possible with the advent of induced pluripotent stem cells (iPSC)-reprogrammed from adult somatic cells, that serves as a starting material for generating retinal cells de novo. Here, we describe a detailed procedure for reprogramming peripheral blood mononuclear cells (PBMC) into iPSC using episomal vectors without any physical disruption in the host genome. The lines thus created were tested for sterility, cytogenetic stability, identity, absence of episomal plasmids and further authenticated for pluripotency and tri-lineage differentiation capacity by embryoid body formation and immunocytochemistry. We believe that this feeder-cell free, animal-product free and gene-insertion free protocol would help people to develop and bank patient-specific cell lines for autologous cell therapies for incurable rare diseases.
MeSH term(s)	Animals ; Cell Differentiation/genetics ; Cellular Reprogramming ; Humans ; Induced Pluripotent Stem Cells ; Leukocytes, Mononuclear ; Plasmids/genetics
Language	English
Publishing date	2021-03-26
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/7651_2021_385
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Article ; Online: An improved protocol for generation and characterization of human-induced pluripotent stem cell-derived retinal pigment epithelium cells.

Surendran, Harshini / Soundararajan, Lalitha / Reddy K, Vijay Bhaskar / Subramani, Janavi / Stoddard, Jonathan / Reynaga, Rene / Tschetter, Wayne / Ryals, Renee C / Pal, Rajarshi

STAR protocols

2022 Volume 3, Issue 4, Page(s) 101803

Abstract: We present an optimized protocol for guided differentiation of retinal pigment epithelium (RPE) cells from human-induced pluripotent stem cells (iPSC). De novo-generated RPE cells are mature, polarized, and mimic the cellular and molecular profile of ... ...

Abstract	We present an optimized protocol for guided differentiation of retinal pigment epithelium (RPE) cells from human-induced pluripotent stem cells (iPSC). De novo-generated RPE cells are mature, polarized, and mimic the cellular and molecular profile of primary RPE; they are also suitable for
MeSH term(s)	Humans ; Retinal Pigment Epithelium ; Induced Pluripotent Stem Cells ; Macular Degeneration/therapy ; Cell Differentiation ; Cell- and Tissue-Based Therapy
Language	English
Publishing date	2022-11-04
Publishing country	United States
Document type	Journal Article
ISSN	2666-1667
ISSN (online)	2666-1667
DOI	10.1016/j.xpro.2022.101803
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Article ; Online: Differentiating Human Induced Pluripotent Stem Cells (iPSCs) Into Lung Epithelial Cells.

Surendran, Harshini / Rajamoorthy, Mohanapriya / Pal, Rajarshi

Current protocols in stem cell biology

2019 Volume 49, Issue 1, Page(s) e86

Abstract: Human induced pluripotent stem cells (hiPSCs) not only offer great opportunities for the study of human development but also have tremendous potential for future clinical cell-based therapies. The protocol outlined here is used to differentiate hiPSCs ... ...

Abstract	Human induced pluripotent stem cells (hiPSCs) not only offer great opportunities for the study of human development but also have tremendous potential for future clinical cell-based therapies. The protocol outlined here is used to differentiate hiPSCs into lung epithelial cell types through a process that faithfully recapitulates the stepwise events observed in vivo. From pluripotency, cells are differentiated to a definitive endoderm fate, followed by progression into anteriorized foregut endoderm that has the ability to give rise to both proximal and distal epithelial cells. Furthermore, this methodology allows for the study of lung dysfunction and disease modeling using patient-derived cells, as well as high-throughput pharmacological screening and eventually personalized therapies. Recently we were able to reproduce this protocol using the working cell bank of an hiPSC line made under current Good Manufacturing Practice (cGMP) conditions, a necessary step for the future clinical application of these cells. © 2019 by John Wiley & Sons, Inc.
MeSH term(s)	Cell Differentiation/physiology ; Cellular Reprogramming/physiology ; Endoderm/cytology ; Epithelial Cells/cytology ; Humans ; Induced Pluripotent Stem Cells/cytology ; Lung/cytology
Language	English
Publishing date	2019-04-18
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1938-8969
ISSN (online)	1938-8969
DOI	10.1002/cpsc.86
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Article ; Online: Human Induced Pluripotent Stem Cell-Derived Lung Epithelial System for SARS-CoV-2 Infection Modeling and Its Potential in Drug Repurposing

Surendran, Harshini / Nandakumar, Swapna / Pal, Rajarshi

Stem Cells and Development ; ISSN 1547-3287 1557-8534

2020

Keywords	Developmental Biology ; Cell Biology ; Hematology ; covid19
Language	English
Publisher	Mary Ann Liebert Inc
Publishing country	us
Document type	Article ; Online
DOI	10.1089/scd.2020.0152
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Article ; Online: Neuronal and cardiac toxicity of pharmacological compounds identified through transcriptomic analysis of human pluripotent stem cell-derived embryoid bodies.

Konala, Vijay Bhaskar Reddy / Nandakumar, Swapna / Surendran, Harshini / Datar, Savita / Bhonde, Ramesh / Pal, Rajarshi

Toxicology and applied pharmacology

2021 Volume 433, Page(s) 115792

Abstract: Concurrent with the '3R' principle, the embryonic stem cell test (EST) using mouse embryonic stem cells, developed in 2000, remains the solely accepted in vitro method for embryotoxicity testing. However, the scope and implementation of EST for ... ...

Abstract	Concurrent with the '3R' principle, the embryonic stem cell test (EST) using mouse embryonic stem cells, developed in 2000, remains the solely accepted in vitro method for embryotoxicity testing. However, the scope and implementation of EST for embryotoxicity screening, compliant with regulatory requirements, are limited. This is due to its technical complexity, long testing period, labor-intensive methodology, and limited endpoint data, leading to misclassification of embryotoxic potential. In this study, we used human induced pluripotent stem cell (hiPSC)-derived embryoid bodies (EB) as an in vitro model to investigate the embryotoxic effects of a carefully selected set of pharmacological compounds. Morphology, viability, and differentiation potential were investigated after exposing EBs to folic acid, all-trans-retinoic acid, dexamethasone, and valproic acid for 15 days. The results showed that the compounds differentially repressed cell growth, compromised morphology, and triggered apoptosis in the EBs. Further, transcriptomics was employed to compare subtle temporal changes between treated and untreated cultures. Gene ontology and pathway analysis revealed that dysregulation of a large number of genes strongly correlated with impaired neuroectoderm and cardiac mesoderm formation. This aberrant gene expression pattern was associated with several disorders of the brain like mental retardation, multiple sclerosis, stroke and of the heart like dilated cardiomyopathy, ventricular tachycardia, and ventricular arrhythmia. Lastly, these in vitro findings were validated using in ovo chick embryo model. Taken together, pharmacological compound or drug-induced defective EB development from hiPSCs could potentially be used as a suitable in vitro platform for embryotoxicity screening.
MeSH term(s)	Animals ; Apoptosis/drug effects ; Cell Differentiation/drug effects ; Cell Line ; Cell Lineage ; Chick Embryo ; Dexamethasone/toxicity ; Dose-Response Relationship, Drug ; Embryoid Bodies/drug effects ; Embryoid Bodies/metabolism ; Embryoid Bodies/pathology ; Gene Expression Profiling ; Gene Expression Regulation, Developmental/drug effects ; Humans ; Induced Pluripotent Stem Cells/drug effects ; Induced Pluripotent Stem Cells/metabolism ; Induced Pluripotent Stem Cells/pathology ; Inhibitory Concentration 50 ; Myocytes, Cardiac/drug effects ; Myocytes, Cardiac/metabolism ; Myocytes, Cardiac/pathology ; Neural Stem Cells/drug effects ; Neural Stem Cells/metabolism ; Neural Stem Cells/pathology ; Neurogenesis/drug effects ; Risk Assessment ; Teratogens/toxicity ; Toxicity Tests ; Transcriptome/drug effects ; Tretinoin/toxicity ; Valproic Acid/toxicity
Chemical Substances	Teratogens ; Tretinoin (5688UTC01R) ; Valproic Acid (614OI1Z5WI) ; Dexamethasone (7S5I7G3JQL)
Language	English
Publishing date	2021-11-03
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	204477-8
ISSN	1096-0333 ; 0041-008X
ISSN (online)	1096-0333
ISSN	0041-008X
DOI	10.1016/j.taap.2021.115792
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Article ; Online: SARS-CoV-2 infection of human-induced pluripotent stem cells-derived lung lineage cells evokes inflammatory and chemosensory responses by targeting mitochondrial pathways.

Surendran, Harshini / Kumar, Saurabh / Narasimhaiah, Swathi / Ananthamurthy, Anuradha / Varghese, P S / D'Souza, George A / Medigeshi, Guruprasad / Pal, Rajarshi

Journal of cellular physiology

2022 Volume 237, Issue 7, Page(s) 2913–2928

Abstract: The COVID-19 disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) primarily affects the lung, particularly the proximal airway and distal alveolar cells. NKX2.1+ primordial lung progenitors of the foregut (anterior) endoderm are ...

Abstract	The COVID-19 disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) primarily affects the lung, particularly the proximal airway and distal alveolar cells. NKX2.1+ primordial lung progenitors of the foregut (anterior) endoderm are the developmental precursors to all adult lung epithelial lineages and are postulated to play an important role in viral tropism. Here, we show that SARS-CoV-2 readily infected and replicated in human-induced pluripotent stem cell-derived proximal airway cells, distal alveolar cells, and lung progenitors. In addition to the upregulation of antiviral defense and immune responses, transcriptomics data uncovered a robust epithelial cell-specific response, including perturbation of metabolic processes and disruption in the alveolar maturation program. We also identified spatiotemporal dysregulation of mitochondrial heme oxygenase 1 (HMOX1), which is associated with defense against antioxidant-induced lung injury. Cytokines, such as TNF-α, INF-γ, IL-6, and IL-13, were upregulated in infected cells sparking mitochondrial ROS production and change in electron transport chain complexes. Increased mitochondrial ROS then activated additional proinflammatory cytokines leading to an aberrant cell cycle resulting in apoptosis. Notably, we are the first to report a chemosensory response resulting from SARS-CoV-2 infection similar to that seen in COVID-19 patients. Some of our key findings were validated using COVID-19-affected postmortem lung tissue sections. These results suggest that our in vitro system could serve as a suitable model to investigate the pathogenetic mechanisms of SARS-CoV-2 infection and to discover and test therapeutic drugs against COVID-19 or its consequences.
MeSH term(s)	Adult ; COVID-19/immunology ; COVID-19/pathology ; Cytokines ; Humans ; Induced Pluripotent Stem Cells/pathology ; Induced Pluripotent Stem Cells/virology ; Lung/pathology ; Lung/virology ; Mitochondria/metabolism ; Reactive Oxygen Species ; SARS-CoV-2
Chemical Substances	Cytokines ; Reactive Oxygen Species
Language	English
Publishing date	2022-04-23
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	3116-1
ISSN	1097-4652 ; 0021-9541
ISSN (online)	1097-4652
ISSN	0021-9541
DOI	10.1002/jcp.30755
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Article: Human Induced Pluripotent Stem Cell-Derived Lung Epithelial System for SARS-CoV-2 Infection Modeling and Its Potential in Drug Repurposing

Surendran, Harshini / Nandakumar, Swapna / Pal, Rajarshi

Stem cells dev

Abstract	The lung is the most vulnerable target for the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection, and respiratory failure causing acute respiratory distress syndrome is its foremost outcome. However, the current primary in vitro models in use for SARS-CoV-2 display apparent limitations for modeling such complex human respiratory disease. Although patient cells can directly model the effects of a drug, their availability and capacity for expansion are limited compared with transformed/immortalized cells or tumor-derived cell lines. An additional caveat is that the latter may harbor genetic and metabolic abnormalities making them unsuitable for drug screening. Therefore, it is important to create physiologically relevant human-cell models that can replicate the pathophysiology of SARS-CoV-2, thus facilitating drug testing. In this study, we show preliminary data on how human induced pluripotent stem cells-derived lung epithelial cell system could emerge as a relevant and sensitive platform for modeling SARS-CoV-2 infection and drug screening.
Keywords	covid19
Publisher	WHO
Document type	Article
Note	WHO #Covidence: #739007
Database	COVID19

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Article ; Online: Generation of Transplantable Retinal Pigmented Epithelial (RPE) Cells for Treatment of Age-Related Macular Degeneration (AMD).

Surendran, Harshini / Rathod, Reena J / Pal, Rajarshi

Methods in molecular biology (Clifton, N.J.)

2018 Volume 2045, Page(s) 283–298

Abstract: Age-related macular degeneration (AMD) is the foremost cause of blindness in people over the age of 60 worldwide. Clinically, this disease starts with distortion in central vision eventually leading to legal blindness. Vision loss has a significant ... ...

Abstract	Age-related macular degeneration (AMD) is the foremost cause of blindness in people over the age of 60 worldwide. Clinically, this disease starts with distortion in central vision eventually leading to legal blindness. Vision loss has a significant impact on quality of life and incurs a substantial cost to the economy. Furthermore, AMD is a complex and progressive neurodegenerative disorder that triggers visual impairment due to the loss of retinal pigmented epithelium (RPE) and the light-sensitive photoreceptors that they support, protect and provide nutrition. Currently, there is no curative treatment for the most common form of this disease, i.e., dry AMD. A novel approach to treat AMD involves the transplantation of RPE cells derived from human induced pluripotent stem cells (iPSCs) in the outer retina. These iPSC-derived RPE cells not only show characteristics similar to native RPE but also could replace as well as regenerate damaged pathologic RPE and produce supportive growth factors and cytokines. Several clinical trials are being conducted taking advantage of a variety of cell- and tissue engineering-based approaches. Here, we present a simple, cost effective, and scalable cell-culture model for generation of purified RPE thus providing the foundation for developing an allogeneic cell therapy for AMD.
MeSH term(s)	ADP-Ribosylation Factors/metabolism ; Antibodies/metabolism ; Cell Culture Techniques/methods ; Cell Differentiation/drug effects ; Cells, Cultured ; Collagen/chemistry ; Drug Combinations ; Flow Cytometry ; Fluorescent Antibody Technique ; Humans ; Induced Pluripotent Stem Cells/cytology ; Induced Pluripotent Stem Cells/drug effects ; Induced Pluripotent Stem Cells/metabolism ; Laminin/chemistry ; Macular Degeneration/therapy ; Nestin/metabolism ; Octamer Transcription Factor-3/metabolism ; PAX6 Transcription Factor/metabolism ; Polymerase Chain Reaction ; Proteoglycans/chemistry ; Retinal Pigment Epithelium/cytology ; Retinal Pigment Epithelium/metabolism ; Retinal Pigment Epithelium/transplantation ; Transcription Factor Brn-3A/metabolism ; Workflow ; cis-trans-Isomerases/metabolism
Chemical Substances	Antibodies ; Drug Combinations ; Laminin ; NES protein, human ; Nestin ; Octamer Transcription Factor-3 ; PAX6 Transcription Factor ; POU4F1 protein, human ; POU5F1 protein, human ; Proteoglycans ; Transcription Factor Brn-3A ; matrigel (119978-18-6) ; Collagen (9007-34-5) ; retinoid isomerohydrolase (EC 3.1.1.64) ; ADP-Ribosylation Factors (EC 3.6.5.2) ; ARL13B protein, human (EC 3.6.5.2) ; cis-trans-Isomerases (EC 5.2.-)
Language	English
Publishing date	2018-06-13
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/7651_2018_140
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Article ; Online: Induced pluripotent stem cells (iPSC)-derived retinal cells in disease modeling and regenerative medicine.

Rathod, Reena / Surendran, Harshini / Battu, Rajani / Desai, Jogin / Pal, Rajarshi

Journal of chemical neuroanatomy

2018 Volume 95, Page(s) 81–88

Abstract: Retinal degenerative disorders are a leading cause of the inherited, irreversible and incurable vision loss. While various rodent model systems have provided crucial information in this direction, lack of disease-relevant tissue availability and species- ... ...

Abstract	Retinal degenerative disorders are a leading cause of the inherited, irreversible and incurable vision loss. While various rodent model systems have provided crucial information in this direction, lack of disease-relevant tissue availability and species-specific differences have proven to be a major roadblock. Human induced pluripotent stem cells (iPSC) have opened up a whole new avenue of possibilities not just in understanding the disease mechanism but also potential therapeutic approaches towards a cure. In this review, we have summarized recent advances in the methods of deriving retinal cell types from iPSCs which can serve as a renewable source of disease-relevant cell population for basic as well as translational studies. We also provide an overview of the ongoing efforts towards developing a suitable in vitro model for modeling retinal degenerative diseases. This basic understanding in turn has contributed to advances in translational goals such as drug screening and cell-replacement therapies. Furthermore we discuss gene editing approaches for autologous repair of genetic disorders and allogeneic transplantation of stem cell-based retinal derivatives for degenerative disorders with an ultimate goal to restore vision. It is pertinent to note however, that these exciting new developments throw up several challenges that need to be overcome before their full clinical potential can be realized.
MeSH term(s)	Animals ; Cell Transplantation ; Humans ; Induced Pluripotent Stem Cells ; Models, Biological ; Regenerative Medicine/methods ; Retina ; Retinal Degeneration
Language	English
Publishing date	2018-02-12
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	639443-7
ISSN	1873-6300 ; 0891-0618
ISSN (online)	1873-6300
ISSN	0891-0618
DOI	10.1016/j.jchemneu.2018.02.002
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