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Drug Screening or Drug Designing Based on Stem Cell Models

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A special issue of Pharmaceuticals (ISSN 1424-8247). This special issue belongs to the section "Biopharmaceuticals".

Deadline for manuscript submissions: closed (31 March 2022) | Viewed by 46490

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Dr. Saima Imran

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Department of Cardiac Thoracic and Vascular Sciences and Public Health, Italy LifeLab Torre della Ricerca, 5 piano sud , Corso Stati Uniti 4, 35127 Padua, Italy
Interests: Human induced pluripotent stem cells; disease modeling; organ regeneration; drug discovery; toxicity screening
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Stem cells have gained great attention in the field of drug discovery and regenerative medicine in recent years. The main advantage of technology, the ability to produce an identical copy of itself, and multipotency, i.e., the ability to generate cells different from itself, are fundamental characteristics of a stem cell. These are unique qualities that have allowed application in various diseases/disorders and medical challenges in both academia and industry—for instance, Parkinson’s disease, Alzheimer’s disease, diabetes, multiple sclerosis, heart disease, cancer, spinal cord injury, wound healing, and organ transplantation.

The prospect of performing high-throughput drug screening is aimed at proliferation, directed differentiation, and toxicity and efficacy studies using stem cells. This seems to be a reliable platform for the drug discovery process. The discovery of induced pluripotent stem cells (hiPSCs) derived from normal or diseased tissue (human and animal) serves as a platform to perform drug screening aimed at developing cell-based therapies against conditions like Parkinson’s disease and diabetes.

There are various research areas in which stem cell technology could make substantial contributions to the development and implementation of stem cell-based models for toxicity testing. Increased use of human in vitro models of toxicity could reduce the use of animals in safety and risk assessment studies and offers the potential to dramatically enhance our understanding of the molecular basis of toxicity, leading to improved human models and assays for predicting biological response to drugs and environmental hazards.

This Special Issue will be providing an opportunity to discuss the application of stem cells and differentiated tissue specific cells from healthy and diseased sources in drug screening and their role in complementing, reducing, and replacing animal trials. In addition to this, target identification and major advances in the field of personalized medicine using induced pluripotent cells will also be focused on.

Dr. Saima Imran
Guest Editor

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Published Papers (12 papers)

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Research

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18 pages, 4019 KiB

Open AccessArticle

Aryl Hydrocarbon Receptor (AhR)-Mediated Signaling in iPSC-Derived Human Motor Neurons

by Saima Jalil Imran, Barbora Vagaska, Jan Kriska, Miroslava Anderova, Mario Bortolozzi, Gino Gerosa, Patrizia Ferretti and Radim Vrzal

Pharmaceuticals 2022, 15(7), 828; https://doi.org/10.3390/ph15070828 - 4 Jul 2022

Cited by 3 | Viewed by 2675

Abstract

Exposure to environmental pollutants and endogenous metabolites that induce aryl hydrocarbon receptor (AhR) expression has been suggested to affect cognitive development and, particularly in boys, also motor function. As current knowledge is based on epidemiological and animal studies, in vitro models are needed to better understand the effects of these compounds in the human nervous system at the molecular level. Here, we investigated expression of AhR pathway components and how they are regulated by AhR ligands in human motor neurons. Motor neurons generated from human induced pluripotent stem cells (hiPSCs) were characterized at the molecular level and by electrophysiology. mRNA levels of AhR target genes, CYP1A1 and CYP1B1 (cytochromes P450 1A1/1B1), and AhR signaling components were monitored in hiPSCs and in differentiated neurons following treatment with AhR ligands, 2,3,7,8,-tetrachlodibenzo-p-dioxin (TCDD), L-kynurenine (L-Kyn), and kynurenic acid (KA), by RT-qPCR. Changes in AhR cellular localization and CYP1A1 activity in neurons treated with AhR ligands were also assessed. The neurons we generated express motor neuron-specific markers and are functional. Transcript levels of CYP1B1, AhR nuclear translocators (ARNT1 and ARNT2) and the AhR repressor (AhRR) change with neuronal differentiation, being significantly higher in neurons than hiPSCs. In contrast, CYP1A1 and AhR transcript levels are slightly lower in neurons than in hiPSCs. The response to TCDD treatment differs in hiPSCs and neurons, with only the latter showing significant CYP1A1 up-regulation. In contrast, TCDD slightly up-regulates CYP1B1 mRNA in hiPSCs, but downregulates it in neurons. Comparison of the effects of different AhR ligands on AhR and some of its target genes in neurons shows that L-Kyn and KA, but not TCDD, regulate AhR expression and differently affect CYP1A1 and CYP1B1 expression. Finally, although TCDD does not significantly affect AhR transcript levels, it induces AhR protein translocation to the nucleus and increases CYP1A1 activity. This is in contrast to L-Kyn and KA, which either do not affect or reduce, respectively, CYP1A1 activity. Expression of components of the AhR signaling pathway are regulated with neuronal differentiation and are differently affected by TCDD, suggesting that pluripotent stem cells might be less sensitive to this toxin than neurons. Crucially, AhR signaling is affected differently by TCDD and other AhR ligands in human motor neurons, suggesting that they can provide a valuable tool for assessing the impact of environmental pollutants. Full article

(This article belongs to the Special Issue Drug Screening or Drug Designing Based on Stem Cell Models)

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16 pages, 6405 KiB

Open AccessArticle

Nobiletin Ameliorates Cellular Damage and Stress Response and Restores Neuronal Identity Altered by Sodium Arsenate Exposure in Human iPSCs-Derived hNPCs

by Sadaf Jahan, Uzair Ahmad Ansari, Arif Jamal Siddiqui, Danish Iqbal, Johra Khan, Saeed Banawas, Bader Alshehri, Mohammed Merae Alshahrani, Suliman A. Alsagaby, Neeru Singh Redhu and Aditya Bhushan Pant

Pharmaceuticals 2022, 15(5), 593; https://doi.org/10.3390/ph15050593 - 12 May 2022

Cited by 15 | Viewed by 3293

Abstract

Environmental exposure to arsenic has been profoundly associated with chronic systemic disorders, such as neurodegeneration, in both experimental models and clinical studies. The neuronal cells of the brain and the nervous system have a limited regeneration capacity, thus making them more vulnerable to exposure to xenobiotics, leading to long-lasting disabilities. The functional and anatomical complexity of these cells hinders the complete understanding of the mechanisms of neurodegeneration and neuroprotection. The present investigations aimed to evaluate the neuroprotective efficacy of a herbal formulation of Nobiletin (NOB) against the toxic insult induced by sodium arsenate (NA) in human neural progenitor cells (hNPCs) derived from human induced pluripotent stem cells (hiPSCs). Prior to the neuroprotective experiments, biologically safe doses of both NOB and NA were ascertained using standard endpoints of cytotoxicity. Thereafter, the hNPCs were exposed to either NOB (50 μM) or NA (50 μM) and co-exposed to biologically safe concentrations of NA (50 μM) with NOB (50 μM) for a period of up to 48 h. NOB treatment restored the morphological damage (neurite damage), the levels of stress granule G3BP1 (Ras-GTPase-activating protein (SH3 domain)-binding protein) and TIA1 (T cell-restricted intracellular antigen), and the expression of neuronal markers (Tuj1, Nestin, MAP2, and PAX6) when compared to NA-exposed cells. A substantial restoration of reactive oxygen species and mitochondrial membrane potential was also witnessed in the co-exposure group (NA + NOB) in comparison to the NA-exposed group. The findings suggest that NOB possesses a significant restorative/protective potential against the NA challenge in hNPCs under experimental conditions and imply that nobiletin may impart a potential therapeutic impact if studied adequately using in vivo studies. Full article

(This article belongs to the Special Issue Drug Screening or Drug Designing Based on Stem Cell Models)

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24 pages, 9469 KiB

Open AccessArticle

Synergistic Adverse Effects of Azithromycin and Hydroxychloroquine on Human Cardiomyocytes at a Clinically Relevant Treatment Duration

by Wener Li, Xiaojing Luo, Mareike S. Poetsch, Reinhard Oertel, Kapil Nichani, Martin Schneider, Anna Strano, Marcel Hasse, Robert-Patrick Steiner, Lukas Cyganek, Karina Hettwer, Steffen Uhlig, Kirsten Simon, Kaomei Guan and Mario Schubert

Pharmaceuticals 2022, 15(2), 220; https://doi.org/10.3390/ph15020220 - 12 Feb 2022

Cited by 4 | Viewed by 3713

Abstract

Adverse effects of drug combinations and their underlying mechanisms are highly relevant for safety evaluation, but often not fully studied. Hydroxychloroquine (HCQ) and azithromycin (AZM) were used as a combination therapy in the treatment of COVID-19 patients at the beginning of the pandemic, leading to higher complication rates in comparison to respective monotherapies. Here, we used human-induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) to systematically investigate the effects of HCQ, AZM, and their combination on the structure and functionality of cardiomyocytes, and to better understand the underlying mechanisms. Our results demonstrate synergistic adverse effects of AZM and HCQ on electrophysiological and contractile function of iPSC-CMs. HCQ-induced prolongation of field potential duration (FPDc) was gradually increased during 7-day treatment period and was strongly enhanced by combination with AZM, although AZM alone slightly shortened FPDc in iPSC-CMs. Combined treatment with AZM and HCQ leads to higher cardiotoxicity, more severe structural disarrangement, more pronounced contractile dysfunctions, and more elevated conduction velocity, compared to respective monotreatments. Mechanistic insights underlying the synergistic effects of AZM and HCQ on iPSC-CM functionality are provided based on increased cellular accumulation of HCQ and AZM as well as increased Cx43- and Nav1.5-protein levels. Full article

(This article belongs to the Special Issue Drug Screening or Drug Designing Based on Stem Cell Models)

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18 pages, 2806 KiB

Open AccessArticle

Testing Mitochondrial-Targeted Drugs in iPSC-RPE from Patients with Age-Related Macular Degeneration

by Mara C. Ebeling, Zhaohui Geng, Madilyn R. Stahl, Rebecca J. Kapphahn, Heidi Roehrich, Sandra R. Montezuma, Deborah A. Ferrington and James R. Dutton

Pharmaceuticals 2022, 15(1), 62; https://doi.org/10.3390/ph15010062 - 4 Jan 2022

Cited by 13 | Viewed by 3398

Abstract

Age-related macular degeneration (AMD) is the leading cause of blindness in the elderly. No universally effective treatments exist for atrophic or “dry” AMD, which results from loss of the retinal pigment epithelium (RPE) and photoreceptors and accounts for ≈80% of all AMD patients. Prior studies provide evidence for the involvement of mitochondrial dysfunction in AMD pathology. This study used induced pluripotent stem cell (iPSC) RPE derived from five AMD patients to test the efficacy of three drugs (AICAR (5-Aminoimidazole-4-carboxamide ribonucleotide), Metformin, trehalose) that target key processes in maintaining optimal mitochondrial function. The patient iPSC-RPE lines were used in a proof-of-concept drug screen, utilizing an analysis of RPE mitochondrial function following acute and extended drug exposure. Results show considerable variability in drug response across patient cell lines, supporting the need for a personalized medicine approach for treating AMD. Furthermore, our results demonstrate the feasibility of using iPSC-RPE from AMD patients to develop a personalized drug treatment regime and provide a roadmap for the future clinical management of AMD. Full article

(This article belongs to the Special Issue Drug Screening or Drug Designing Based on Stem Cell Models)

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9 pages, 544 KiB

Open AccessCommunication

Testing the Stability of Drug Resistance on Cryopreserved, Gene-Engineered Human Induced Pluripotent Stem Cells

by Dilaware Khan, Ann-Christin Nickel, Sebastian Jeising, Constanze Uhlmann, Sajjad Muhammad, Daniel Hänggi, Igor Fischer and Ulf Dietrich Kahlert

Pharmaceuticals 2021, 14(9), 919; https://doi.org/10.3390/ph14090919 - 11 Sep 2021

Cited by 1 | Viewed by 2535

Abstract

Human induced pluripotent stem cells (hiPSCs) have emerged as a powerful tool for in vitro modelling of diseases with broad application in drug development or toxicology testing. These assays usually require large quantities of hiPSC, which can entail long-term storage via cryopreservation of the same cell charges. However, it is essential that cryopreservation does not oppose durable changes on the cells. In this project, we characterize one parameter of functionality of one that is well established in the field, in a different research context, an applied hiPSC line (iPS11), namely their resistance to a medium size library of chemo interventions (>160 drugs). We demonstrate that cells, before and after cryopreservation, do not change their relative overall drug response phenotypes, as defined by identification of the top 20 interventions causing dose-dependent reduction of cell growth. Importantly, also frozen cells that are exogenously enforced for stable overexpression of oncogenes myelocytomatosis (cMYC) or tumor protein 53 mutation (TP53R175H), respectively, are not changed in their relative top 20 drugs response compared to their non-frozen counterparts. Taken together, our results support iPSCs as a reliable in vitro platform for in vitro pharmacology, further raising hopes that this technology supports biomarker-associated drug development. Given the general debate on ethical and economic problems associated with the reproducibly crisis in biomedicine, our results may be of interest to a wider audience beyond stem cell research. Full article

(This article belongs to the Special Issue Drug Screening or Drug Designing Based on Stem Cell Models)

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15 pages, 4445 KiB

Open AccessArticle

Thyroid Hormone Effect on the Differentiation of Human Induced Pluripotent Stem Cells into Hepatocyte-Like Cells

by Mariia S. Bogacheva, Margarita A. Bystriakova and Yan-Ru Lou

Pharmaceuticals 2021, 14(6), 544; https://doi.org/10.3390/ph14060544 - 7 Jun 2021

Cited by 4 | Viewed by 3241

Abstract

Human induced pluripotent stem cells (hiPSCs) hold great potential as an unlimited source for obtaining hepatocyte-like cells (HLCs) for drug research. However, current applications of HLCs have been severely limited by the inability to produce mature hepatocytes from hiPSCs in vitro. Thyroid hormones are one of the hormones that surge during the perinatal period when liver maturation takes place. Here we assessed the influence of thyroid hormone on hepatic progenitor differentiation to HLCs. We analyzed gene and protein expression of early and late hepatic markers and demonstrated the selective activity of thyroid hormone on different genes. Particularly, we demonstrated thyroid hormone-dependent inhibition of the fetal hepatic marker AFP. Our study sheds light on the role of thyroid hormone during liver differentiation and maturation. Full article

(This article belongs to the Special Issue Drug Screening or Drug Designing Based on Stem Cell Models)

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Review

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16 pages, 1866 KiB

Open AccessReview

Induced Pluripotent Stem Cell-Based Drug Screening by Use of Artificial Intelligence

by Dai Kusumoto, Shinsuke Yuasa and Keiichi Fukuda

Pharmaceuticals 2022, 15(5), 562; https://doi.org/10.3390/ph15050562 - 30 Apr 2022

Cited by 15 | Viewed by 5801

Abstract

Induced pluripotent stem cells (iPSCs) are terminally differentiated somatic cells that differentiate into various cell types. iPSCs are expected to be used for disease modeling and for developing novel treatments because differentiated cells from iPSCs can recapitulate the cellular pathology of patients with genetic mutations. However, a barrier to using iPSCs for comprehensive drug screening is the difficulty of evaluating their pathophysiology. Recently, the accuracy of image analysis has dramatically improved with the development of artificial intelligence (AI) technology. In the field of cell biology, it has become possible to estimate cell types and states by examining cellular morphology obtained from simple microscopic images. AI can evaluate disease-specific phenotypes of iPS-derived cells from label-free microscopic images; thus, AI can be utilized for disease-specific drug screening using iPSCs. In addition to image analysis, various AI-based methods can be applied to drug development, including phenotype prediction by analyzing genomic data and virtual screening by analyzing structural formulas and protein–protein interactions of compounds. In the future, combining AI methods may rapidly accelerate drug discovery using iPSCs. In this review, we explain the details of AI technology and the application of AI for iPSC-based drug screening. Full article

(This article belongs to the Special Issue Drug Screening or Drug Designing Based on Stem Cell Models)

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22 pages, 1249 KiB

Open AccessReview

Human Pluripotent Stem-Cell-Derived Models as a Missing Link in Drug Discovery and Development

by Xiying Lin, Jiayu Tang and Yan-Ru Lou

Pharmaceuticals 2021, 14(6), 525; https://doi.org/10.3390/ph14060525 - 30 May 2021

Cited by 13 | Viewed by 4971

Abstract

Human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) and human-induced pluripotent stem cells (hiPSCs), have the potential to accelerate the drug discovery and development process. In this review, by analyzing each stage of the drug discovery and development process, we identified the active role of hPSC-derived in vitro models in phenotypic screening, target-based screening, target validation, toxicology evaluation, precision medicine, clinical trial in a dish, and post-clinical studies. Patient-derived or genome-edited PSCs can generate valid in vitro models for dissecting disease mechanisms, discovering novel drug targets, screening drug candidates, and preclinically and post-clinically evaluating drug safety and efficacy. With the advances in modern biotechnologies and developmental biology, hPSC-derived in vitro models will hopefully improve the cost-effectiveness and the success rate of drug discovery and development. Full article

(This article belongs to the Special Issue Drug Screening or Drug Designing Based on Stem Cell Models)

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27 pages, 2021 KiB

Open AccessReview

Examining the Characteristics and Applications of Mesenchymal, Induced Pluripotent, and Embryonic Stem Cells for Tissue Engineering Approaches across the Germ Layers

by Caitlin Priester, Amber MacDonald, Madhu Dhar and Austin Bow

Pharmaceuticals 2020, 13(11), 344; https://doi.org/10.3390/ph13110344 - 26 Oct 2020

Cited by 18 | Viewed by 3567

Abstract

The field of regenerative medicine utilizes a wide array of technologies and techniques for repairing and restoring function to damaged tissues. Among these, stem cells offer one of the most potent and promising biological tools to facilitate such goals. Implementation of mesenchymal stem cells (MSCs), induced pluripotent stem cells (iPSCs), and embryonic stem cells (ESCs) offer varying advantages based on availability and efficacy in the target tissue. The focus of this review is to discuss characteristics of these three subset stem cell populations and examine their utility in tissue engineering. In particular, the development of therapeutics that utilize cell-based approaches, divided by germinal layer to further assess research targeting specific tissues of the mesoderm, ectoderm, and endoderm. The combinatorial application of MSCs, iPSCs, and ESCs with natural and synthetic scaffold technologies can enhance the reparative capacity and survival of implanted cells. Continued efforts to generate more standardized approaches for these cells may provide improved study-to-study variations on implementation, thereby increasing the clinical translatability of cell-based therapeutics. Coupling clinically translatable research with commercially oriented methods offers the potential to drastically advance medical treatments for multiple diseases and injuries, improving the quality of life for many individuals. Full article

(This article belongs to the Special Issue Drug Screening or Drug Designing Based on Stem Cell Models)

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8 pages, 235 KiB

Open AccessCommentary

Allogenic Amniotic Tissue for Treatment of Knee and Hip Osteoarthritis

by Ashim Gupta

Pharmaceuticals 2022, 15(4), 404; https://doi.org/10.3390/ph15040404 - 26 Mar 2022

Cited by 12 | Viewed by 2685

Abstract

Osteoarthritis (OA) impacts millions of people and places a high burden on healthcare systems in the United States. Current treatment modalities have limitations and do not address underlying pathology. Lately, there has been an immense growth in the use of biologics, including perinatal allogenic tissues for orthopedic regenerative medicine applications. Amniotic tissue is an exciting new alternative for such applications. Despite several published studies that reported its use for treatment of ophthalmic conditions and complex wounds, there are limited clinical studies evaluating its safety and efficacy in treating patients suffering with knee or hip OA. In this manuscript, I focused on three prospective clinical studies which evaluated the safety and efficacy of amniotic tissue in patients suffering with moderate knee or hip OA. The results from these studies presented the scientific community with much needed, well-executed, and prospective clinical trials. Though these trials demonstrated that administration of amniotic tissue in knee or hip joint is safe and potentially effective, more multi-center, prospective, double-blinded, randomized controlled trials are warranted to further establish the efficacy of amniotic tissue to mitigate symptoms of knee and hip OA to ultimately justify its clinical use. Full article

(This article belongs to the Special Issue Drug Screening or Drug Designing Based on Stem Cell Models)

11 pages, 1050 KiB

Open AccessSystematic Review

Umbilical Cord-Derived Wharton’s Jelly for Regenerative Medicine Applications: A Systematic Review

by Benjamin J. Main, Nicola Maffulli, Josiah A. Valk, Hugo C. Rodriguez, Manu Gupta, Saadiq F. El-Amin III and Ashim Gupta

Pharmaceuticals 2021, 14(11), 1090; https://doi.org/10.3390/ph14111090 - 27 Oct 2021

Cited by 17 | Viewed by 4392

Abstract

Musculoskeletal ailments affect millions of people around the world and place a high burden on healthcare. Traditional treatment modalities are limited and do not address underlying pathologies. Mesenchymal stem cells (MSCs) have emerged as an exciting therapeutic alternative and Wharton’s jelly-derived mesenchymal stem cells (WJSCs) are some of these. This review reports the clinical and functional outcomes of the applications of WJSCs in orthopedic surgery. A systematic review was conducted utilizing the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidelines. The studies that used culture-expanded, mesenchymal stem or stromal cells, MSCs and/or connective tissues procured from Wharton’s jelly (WJ), from January 2010 to October 2021, were included. Conventional non-operative therapies and placebos were used as comparisons. Six studies that directly discussed WJSCs use in an animal model or the basic scientific testing using an injury model were identified. Five publications studied cartilage injury, three studied degenerative disc disease, one was related to osteoarthritis, and one was related to osteochondral defects. The results of these studies suggested the benefits of WJSCs in the management of these orthopedic pathologies. To adequately assess the safety and efficacy of WJSCs in orthopedic surgery, further randomized controlled clinical studies are necessary. Full article

(This article belongs to the Special Issue Drug Screening or Drug Designing Based on Stem Cell Models)

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7 pages, 1979 KiB

Open AccessCase Report

Treatment of Knee Osteoarthritis with Intraarticular Umbilical Cord-Derived Wharton’s Jelly: A Case Report

by Ashim Gupta, Hugo C. Rodriguez, Anish G. Potty, Howard J. Levy and Saadiq F. El-Amin III

Pharmaceuticals 2021, 14(9), 883; https://doi.org/10.3390/ph14090883 - 31 Aug 2021

Cited by 9 | Viewed by 4147

Abstract

We present the case of a 27-year-old male with grade II knee osteoarthritis (OA) that was intraarticularly injected with a 2 mL umbilical cord-derived Wharton’s jelly (UC-derived WJ) formulation. The patients’ baseline radiographs were taken and baseline numeric pain rating scale (NPRS), knee injury and osteoarthritis outcome score (KOOS), 7-point Likert scale, and a 36-item short form survey (SF-36) were recorded. The NPRS was re-recorded immediately after the injection, and at 24 h, 48 h, 1 week, 6 weeks, and at 3 months follow-up post-injection. The KOOS and 7-point Likert scale was re-recorded at the patients’ 1week, 6 week, and 3month follow-up, and SF-36 was re-recorded at 3 months. A final set of X-rays were also performed at 3 months follow-up post-injection. No adverse effects from the injection were reported over the duration of the study. No significant difference nor progression in OA on X-rays compared to baseline was observed. NPRS decreased by 50% and the 7-point Likert scale increased to Extremely Satisfied. KOOS increased overall by 10% and the SF-36 overall change was 25%. These results indicate the potential application of UC-derived WJ in the treatment of knee OA. Larger, long term, non-randomized and randomized control trials are warranted to adequately assess the safety and efficacy of UC-derived WJ and ultimate clinical use. Full article

(This article belongs to the Special Issue Drug Screening or Drug Designing Based on Stem Cell Models)

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