Recent Advances in iPSCs for Therapy

The series Advances in Stem Cell Biology is a timely and expansive collection of comprehensive information and new discoveries in the field of stem cell biology. Recent Advances in iPSCs for Therapy, Volume 3 addresses the use of induced pluripotent stem cells for therapy of several disorders.

The volume teaches the reader about the biology of induced pluripotent stem cells and their possible use as cell therapy. Remarkable progress has been made in the obtention of induced pluripotent stem cells and their differentiation into several cell types, tissues and organs using state-of-art techniques.

This volume will cover what we know so far about the use of iPSCs for therapy of multiple diseases, such as: hair loss disorders, Parkinson’s disease, Huntington’s disease, ischemic stroke, Alzheimer’s disease, Glaucoma, Optic Neuropathy, Age-Related Macular Degeneration, Type 1 Diabetes, Heart Failure, liver diseases, infertility, Autoimmune Diseases, and more.

The volume is written for researchers and scientists interested in cell therapy, stem cell biology, regenerative medicine, and organ transplantation and is contributed by world-renowned authors in the field.

• Provides overview of the fast-moving field of stem cell biology and function, regenerative medicine, and therapeutics
• Covers the possible use of iPSCs for hair loss disorders, Parkinson’s disease, Huntington’s disease, ischemic stroke, Alzheimer’s disease, Glaucoma, Optic Neuropathy, Age-Related Macular Degeneration, Type 1 Diabetes, Heart Failure, liver diseases, infertility, Autoimmune Diseases, and more
• Contributions from stem cell leaders around the world

• Language: English
• Publisher: Academic Press
• Release date: Jan 30, 2021
• ISBN: 9780323851862

Book preview

Recent Advances in iPSCs for Therapy, Volume 3

Editor

Alexander Birbrair

Federal University of Minas Gerais, Department of Pathology, Belo Horizonte, Minas Gerais, Brazil

Columbia University Medical Center, Department of Radiology, New York, NY, United States

Table of Contents

Cover image

Title page

Copyright

Dedication

Contributors

About the editor

Preface

Chapter 1. Strategies to utilize iPS cells for hair follicle regeneration and the treatment of hair loss disorders

1. Introduction

2. HF morphology and physiology

3. Experimental approaches to HF regeneration

4. Relationship between HF and iPSCs: the HF as a favorable iPSC material

5. Regeneration of human HFs using hiPSCs

6. Strategies for treating hair loss disorders using hiPSCs

7. Conclusion

List of abbreviations

Chapter 2. iPSCs and cell therapy for Parkinson’s disease

1. Introduction

2. PD as an established target for cell therapy

3. Developing a cell therapy approach for PD

4. Clinical factors

5. Toward clinical implementation of iPSC-based therapy for PD

6. Perspective

Abbreviations

Chapter 3. Induced pluripotent stem cells as a potential treatment for Huntington’s disease

1. Huntington’s disease

2. HTT gene and protein function

3. Epigenetics and HD

4. Signs and symptoms of HD

5. Stem cell therapy for HD

6. Induced pluripotent stem cells (iPSCs)

7. Stem cells–based HD clinical trials

8. Summary and conclusions

Chapter 4. Present and future of adult stem cells and induced pluripotent stem cells therapy for ischemic stroke

1. Introduction

2. Protective and recovery approaches for ischemic stroke

3. Adult stem cells: NSCs and mesenchymal stem cells

4. Induced pluripotent stem cells: a multifaceted role in the study of neurological diseases

5. Conclusion and future perspectives

Glossary

Chapter 5. Stem cell therapy in Alzheimer’s disease

1. Introduction

2. Preclinical studies on the use of stem cells in dementias

3. Endogenous approach

4. Exogenous approach

5. Previous human experience with MSCs

Chapter 6. Stem cell therapies for glaucoma and optic neuropathy

1. Introduction

2. Retinal cell fate specification

3. Retinal cell transplantation

4. iPSCs for modeling of familial glaucoma

5. Summary and future directions

Chapter 7. Induced Pluripotent Stem Cells (iPSC) in Age-related Macular Degeneration (AMD)

1. Background

2. Cellular Therapies for AMD

3. Future Directions

Chapter 8. Considerations in using human pluripotent stem cell–derived pancreatic beta cells to treat type 1 diabetes

1. Introduction

2. hPSC-based clinical trials

3. Considerations for hPSCs for clinical trials

4. hiPSC biobanking efforts

5. Generation of hPSCs

6. Manufacturing hPSC-derived pancreatic beta cells for cell therapy

7. hPSC-based therapy for type 1 diabetes

8. Future outlook and concluding thoughts

Chapter 9. Induced pluripotent stem cells for treatment of heart failure

1. Introduction

2. Trend of cell therapy

3. Production of iPSCs and establishment of a cardiogenic differentiation method

4. Large-scale cell culture for cell transplantation therapy

5. Proof of concept in iPSC-CM sheet for heart failure and immunologic study of iPSCs

6. Removal of undifferentiated iPSCs to confirm safety for clinical applications

7. Development of new drug-based heart failure therapy using disease-specific iPSCs

Chapter 10. Induced pluripotent stem cells in liver disease

1. Introduction

2. Generation of iPSCs and differentiation into hepatic phenotype

3. Use of iPSCs in liver disease

4. iPSC for modeling liver disease

5. Hepatotoxicity studies

6. Conclusions

Abbreviations

Chapter 11. Induced pluripotent stem cells: potential therapeutic application for improving fertility in humans and animals

1. Biology of iPSC in mammals

2. Stem cells as candidates for in vitro germ cell derivation

3. Potential strategies for the use of iPSC-derived germ cells in human, livestock, and wild animal reproductive biotechnology

4. Paracrine control and gene signaling pathways involved in in vivo male germ differentiation: potential approaches for in vitro derivation of germ cells from iPSC

5. In vitro approaches for derivation of germ cells from iPSCs

6. Conclusion

Chapter 12. Induced pluripotent stem cells in wound healing

1. Introduction

2. Background

3. Stem cell therapy in wound healing

4. Induced pluripotent stem cells

5. Challenges and solutions of iPSC in wound healing

6. Conclusion

7. Future directions

Chapter 13. Induced pluripotent stem cells for periodontal regeneration

1. Prevalence of periodontal diseases

2. Overview of pathogenesis of periodontitis

3. Current therapies for periodontitis and their limitations

4. Development of the dental complex

5. Stem cells and tissue regeneration

6. iPSCs in regenerative dentistry

7. Clinical application of iPSCs

8. Conclusions and perspective

Chapter 14. Current development in iPSC-based therapy for autoimmune diseases

1. Introduction

2. Cellular components in autoimmunity

3. Treg cells in autoimmune disease

4. Dendritic cells (DCs) in autoimmune disease

5. Autoimmune disease

6. Diabetes mellitus

7. Rheumatoid arthritis (RA)

8. Multiple sclerosis (MS)

9. Conclusion and future challenges

Index

Copyright

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ISBN: 978-0-12-822229-4

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Dedication

This book is dedicated to my mother, Marina Sobolevsky, of blessed memory, who passed away during the creation of this volume. Professor of Mathematics at the State University of Ceará (UECE), she was loved by her colleagues and students, whom she inspired by her unique manner of teaching. All success in my career and personal life I owe to her.

My beloved father, Lev Birbrair, and my beloved mom, Marina Sobolevsky, of blessed memory (July 28, 1959–June 3, 2020)

Contributors

Graham Anderson, PhD ,     Centre for Regenerative Medicine, Institute for Regeneration and Repair, The University of Edinburgh, Edinburgh BioQuarter, Edinburgh, United Kingdom

Pierre Bagnaninchi, PhD ,     Centre for Regenerative Medicine, Institute for Regeneration and Repair, The University of Edinburgh, Edinburgh BioQuarter, Edinburgh, United Kingdom

Bernard Baumel, MD ,     Department of Neurology, University of Miami Miller School of Medicine, Miami, FL, United States

Alexander Birbrair

Department of Pathology, Federal University of Minas Gerais, Belo Horizonte, MG, Brazil

Department of Radiology, Columbia University Medical Center, New York, NY, United States

Ryan Bloomquist,     Department of Restorative Sciences, The Dental College of Georgia, Augusta University, Augusta, GA, United States

Ana Bugallo-Casal,     Clinical Neurosciences Research Laboratory (LINC), Health Research Institute of Santiago de Compostela (IDIS), Santiago de Compostela, A Coruña, Spain

Christian Camargo, MD ,     Department of Neurology, University of Miami Miller School of Medicine, Miami, FL, United States

Francisco Campos,     Clinical Neurosciences Research Laboratory (LINC), Health Research Institute of Santiago de Compostela (IDIS), Santiago de Compostela, A Coruña, Spain

Kun-Che Chang

Spencer Center for Vision Research, School of Medicine, Stanford University, Palo Alto, CA, United States

Department of Ophthalmology, Louis J. Fox Center for Vision Restoration, University of Pittsburgh School of Medicine, Pittsburgh, PA, United States

Umber Cheema,     UCL Institute of Orthopaedics & Musculoskeletal Sciences, UCL Division of Surgery & Interventional Sciences, University College London, London, United Kingdom

Alan Dardik,     Vascular Biology and Therapeutics Program and the Department of Surgery, Yale University School of Medicine, New Haven, CT, United States

Jugal Kishore Das,     Department of Microbial Pathogenesis and Immunology, Texas A&M University Health Science Center, Bryan, TX, United States

Baljean Dhillon, BMedSci(Hons), BMBS FRCS(Glasg), FRCOphth, FRCS(Ed), FRCPE

Centre for Clinical Brain Sciences, University of Edinburgh, Edinburgh Bioquarter, Edinburgh, United Kingdom

NHS Lothian, Clinical Ophthalmology, Princess Alexandria Eye Pavilion, Edinburgh, United Kingdom

M. Teresa Donato

Unidad de Hepatología Experimental, Instituto de Investigación Sanitaria La Fe, Valencia, Spain

Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad de Valencia, Valencia, Spain

G.L. Dunbar

Field Neurosciences Institute Laboratory for Restorative Neurology, Central Michigan University, Mt. Pleasant, MI, United States

Program in Neuroscience, Central Michigan University, Mt. Pleasant, MI, United States

Department of Psychology, Central Michigan University, Mt. Pleasant, MI, United States

Field Neurosciences Institute, St. Mary’s of Michigan, Saginaw, MI, United States

Xixiang Gao

Department of Vascular Surgery, Xuanwu Hospital, Capital Medical University and Institute of Vascular Surgery, Capital Medical University, Beijing, China

Vascular Biology and Therapeutics Program and the Department of Surgery, Yale University School of Medicine, New Haven, CT, United States

Jolanta Gorecka,     Vascular Biology and Therapeutics Program and the Department of Surgery, Yale University School of Medicine, New Haven, CT, United States

Yongquan Gu,     Department of Vascular Surgery, Xuanwu Hospital, Capital Medical University and Institute of Vascular Surgery, Capital Medical University, Beijing, China

Chin Meng Khoo

Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore

Division of Endocrinology, National University Hospital, Singapore, Singapore

Kwang-Soo Kim

Department of Psychiatry

Molecular Neurobiology Laboratory, McLean Hospital, Harvard Medical School, Belmont, MA, United States

Anil Kumar,     Department of Microbial Pathogenesis and Immunology, Texas A&M University Health Science Center, Bryan, TX, United States

Hwee Hui Lau

Stem Cells and Diabetes Laboratory, Institute of Molecular and Cell Biology, A∗STAR, Singapore, Singapore

School of Biological Sciences, Nanyang Technological University, Singapore, Singapore

Ziming Luo,     Spencer Center for Vision Research, School of Medicine, Stanford University, Palo Alto, CA, United States

Michelle Marrero, MD ,     Department of Neurology, University of Miami Miller School of Medicine, Miami, FL, United States

Shigeru Miyagawa,     Department of Cardiovascular Surgery, Osaka University Graduate School of Medicine, Suita, Osaka, Japan

Mahmood S. Mozaffari,     Department of Oral Biology and Diagnostic Sciences, The Dental College of Georgia, Augusta University, Augusta, GA, United States

Michael Nahmou,     Spencer Center for Vision Research, School of Medicine, Stanford University, Palo Alto, CA, United States

Manabu Ohyama,     Department of Dermatology, Kyorin University Faculty of Medicine, Mitaka-shi, Tokyo, Japan

Víctor H. Parraguez

Department of Biological Sciences, Faculty of Veterinary and Animal Sciences, University of Chile, Santiago, Chile

Department of Animal Production, Faculty of Agrarian Sciences, University of Chile, Santiago, Chile

María Pelechá,     Unidad de Hepatología Experimental, Instituto de Investigación Sanitaria La Fe, Valencia, Spain

Hao-Yun Peng,     Department of Microbial Pathogenesis and Immunology, Texas A&M University Health Science Center, Bryan, TX, United States

Oscar A. Peralta

Department of Animal Production Sciences, Faculty of Veterinary and Animal Sciences, University of Chile, Santiago, Chile

Department of Biomedical Sciences and Pathobiology, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Tech, Blacksburg, VA, United States

Milena Pinto, PhD ,     Department of Neurology, University of Miami Miller School of Medicine, Miami, FL, United States

María Pérez-Mato,     Neuroscience and Cerebrovascular Research Laboratory, Department of Neurology and Stroke Center, La Paz University Hospital, Neuroscience Area of IdiPAZ Health Research Institute, Universidad Autónoma de Madrid, Madrid, Spain

Yijie Ren,     Department of Microbial Pathogenesis and Immunology, Texas A&M University Health Science Center, Bryan, TX, United States

J. Rossignol

College of Medicine, Central Michigan University, Mt. Pleasant, MI, United States

Field Neurosciences Institute Laboratory for Restorative Neurology, Central Michigan University, Mt. Pleasant, MI, United States

Program in Neuroscience, Central Michigan University, Mt. Pleasant, MI, United States

Yoshiki Sawa,     Department of Cardiovascular Surgery, Osaka University Graduate School of Medicine, Suita, Osaka, Japan

Jeffrey S. Schweitzer,     Department of Neurosurgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, United States

Bin Song

Department of Psychiatry

Molecular Neurobiology Laboratory, McLean Hospital, Harvard Medical School, Belmont, MA, United States

Jianxun Song,     Department of Microbial Pathogenesis and Immunology, Texas A&M University Health Science Center, Bryan, TX, United States

B. Srinageshwar

College of Medicine, Central Michigan University, Mt. Pleasant, MI, United States

Field Neurosciences Institute Laboratory for Restorative Neurology, Central Michigan University, Mt. Pleasant, MI, United States

Program in Neuroscience, Central Michigan University, Mt. Pleasant, MI, United States

Wei Xuan Tan

Stem Cells and Diabetes Laboratory, Institute of Molecular and Cell Biology, A∗STAR, Singapore, Singapore

Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore

Nguan Soon Tan

School of Biological Sciences, Nanyang Technological University, Singapore, Singapore

Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore, Singapore

Adrian Kee Keong Teo

Stem Cells and Diabetes Laboratory, Institute of Molecular and Cell Biology, A∗STAR, Singapore, Singapore

Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore

Laia Tolosa,     Unidad de Hepatología Experimental, Instituto de Investigación Sanitaria La Fe, Valencia, Spain

Cristian G. Torres,     Department of Clinical Sciences, Faculty of Veterinary and Animal Sciences, University of Chile, Santiago, Chile

Liqing Wang,     Department of Microbial Pathogenesis and Immunology, Texas A&M University Health Science Center, Bryan, TX, United States

Yingfeng Wu,     Department of Vascular Surgery, Xuanwu Hospital, Capital Medical University and Institute of Vascular Surgery, Capital Medical University, Beijing, China

Xiaofang Xiong,     Department of Microbial Pathogenesis and Immunology, Texas A&M University Health Science Center, Bryan, TX, United States

About the editor

Alexander Birbrair

Dr. Alexander Birbrair received his bachelor's biomedical degree from Santa Cruz State University in Brazil. He completed his PhD in Neuroscience, in the field of stem cell biology, at the Wake Forest School of Medicine under the mentorship of Osvaldo Delbono. Then, he joined as a postdoc in stem cell biology at Paul Frenette's laboratory at Albert Einstein School of Medicine in New York. In 2016, he was appointed faculty at Federal University of Minas Gerais in Brazil, where he started his own lab. His laboratory is interested in understanding how the cellular components of different tissues function and control disease progression. His group explores the roles of specific cell populations in the tissue microenvironment by using state-of-the-art techniques. His research is funded by the Serrapilheira Institute, CNPq, CAPES, and FAPEMIG. In 2018, Alexander was elected affiliate member of the Brazilian Academy of Sciences (ABC), and in 2019, he was elected member of the Global Young Academy (GYA). He is the Founding Editor and Editor-in-Chief of Current Tissue Microenvironment Reports and Associate Editor of Molecular Biotechnology. Alexander also serves in the editorial board of several other international journals: Stem Cell Reviews and Reports, Stem Cell Research, Stem Cells and Development, and Histology and Histopathology.

Preface

Alexander Birbrair ¹ , ² ,      ¹ Department of Pathology, Federal University of Minas Gerais, Belo Horizonte, MG, Brazil,      ² Department of Radiology, Columbia University Medical Center, New York, NY, United States

This book's initial title was iPSCs: Recent Advances. Nevertheless, because of the ongoing strong interest in this theme, we were capable of collecting more chapters than would fit in one single volume, covering induced pluripotent stem cells (iPSCs) biology from different perspectives. Therefore, the book was subdivided into several volumes.

This volume Recent Advances in iPSCs for Therapy offers contributions by known scientists and clinicians in the multidisciplinary areas of biological and medical research. The chapters bring up-to-date comprehensive overviews of current advances in the field. This book describes the use of iPSCs for therapy of several disorders. Further insights into the biology of these cells will have important implications for the possible use of iPSCs as cell therapy. The authors focus on the modern state-of-the-art methodologies and the leading-edge concepts in the field of stem cell biology. In recent years, remarkable progress has been made in the obtention of iPSCs and their differentiation into several cell types, tissues, and organs using state-of-the-art techniques. These advantages facilitated identification of key targets and definition of the molecular basis of several disorders. Thus, the present book is an attempt to describe the most recent developments in the area of iPSCs biology, which is one of the rising hot topics in the field of molecular and cellular biology today. Here, we present a selected collection of detailed chapters on what we know so far about the use of iPSCs for therapy of multiple diseases. Fourteen chapters written by experts in the field summarize the present knowledge about iPSCs therapeutic potential.

Manabu Ohyama from Kyorin University Faculty of Medicine discusses iPSCs for hair follicle regeneration and the treatment of hair loss disorders. Jeffrey S. Schweitzer and colleagues from Harvard Medical School describe iPSCs for Parkinson's disease. Julien Rossignol and colleagues from Central Michigan University compile our understanding of iPSCs as a potential treatment for Huntington's disease. Francisco Campos and colleagues from Health Research Institute of Santiago de Compostela update us with what we know about iPSCs for therapy of ischemic stroke. Barry Baumel and colleagues from University of Miami Miller School of Medicine summarize current knowledge on stem cell therapies for Alzheimer's disease. Kun-Che Chang and colleagues from Stanford University address the importance of iPSCs for glaucoma and optic neuropathy. Graham Anderson and colleagues from University of Edinburgh talk about iPSCs in age-related macular degeneration. Adrian Kee Keong Teo and colleagues from National University of Singapore, Singapore, focus on the use of iPSCs to treat type 1 diabetes. Shigeru Miyagawa and Yoshiki Sawa from Osaka University Graduate School of Medicine give an overview of the use of iPSCs for the treatment of heart failure. Laia Tolosa and colleagues from Universidad de Valencia present iPSCs for liver diseases. Oscar A. Peralta and colleagues from University of Chile introduce what we know so far about potential therapeutic applications of iPSCs for improving fertility in humans and animals. Alan Dardik and colleagues from Yale University School of Medicine discuss iPSCs for wound healing. Mahmood S. Mozaffari and colleagues from Augusta University update us with information on the use of iPSCs for regeneration of dental complex. Finally, Anil Kumar and colleagues from Texas A&M University Health Science Center summarize our current status on the use of iPSCs for autoimmune diseases.

It is hoped that the articles published in this book will become a source of reference and inspiration for future research ideas. I would like to express my deep gratitude to my wife, Veranika Ushakova, and Ms. Billie Jean Fernandez and Ms. Elisabeth Brown from Elsevier, who helped at every step of the execution of this project.

Alexander Birbrair

Editor

Chapter 1: Strategies to utilize iPS cells for hair follicle regeneration and the treatment of hair loss disorders

Manabu Ohyama     Department of Dermatology, Kyorin University Faculty of Medicine, Mitaka-shi, Tokyo, Japan

Abstract

The hair follicle (HF) is a mammalian skin structure that provides physical protection, detects sensation, and enables thermoregulation. In humans, loss of hairs on the head greatly affects the physical appearance, leading to altered quality of life. Thus, vast demand exists for treatments for hair loss disorders, represented by male and female pattern hair loss or alopecia areata. Regenerative medicine approaches, including HF bioengineering, can provide remedies for intractable hair loss diseases caused by irreversible HF destruction. The basis for experimental HF regeneration has been established in murine. However, human HF reconstitution adopting that principle has been hampered by the paucity of starting materials, including HF epithelial stem cells and hair-inductive dermal cells, and the loss of their HF-prone properties during in vitro expansion. With their high-proliferative capacity and multipotency, human induced pluripotent stem cells (hiPSCs) should be useful for HF regeneration. Indeed, hiPSC-derived epithelial and mesenchymal cells can contribute to in vivo HF-like structure regeneration. hiPSCs can also give rise to 3D integumentary organ systems comprising HFs which can be isolated and grafted onto areas of hair loss. Previous studies have focused mainly on the reproduction of HF structures; however, considering that the regeneration of complete HFs is not required to correct many common hair loss conditions, hiPSCs may be better differentiated into trichogenic dermal cells for cell-based therapy. For immune-mediated hair loss disorders, immunoregulatory cells can be induced from hiPSCs and inoculated into the affected lesion. In summary, hiPSCs are a promising cell source not only for HF bioengineering and but also for preparing cell populations that may be able to mitigate hair loss.

Keywords

Alopecia areata; Bulge; Dermal papilla; Dermal sheath; Differentiation; Female pattern hair loss; Hair cycle; Hair follicle; Hair loss; Induced pluripotent stem cells; Keratinocyte; Male pattern hair loss; Mesencymal stem/stromal cells; Primary scarring alopecia; Regeneration; Treatment

Acknowledgment

1. Introduction

2. HF morphology and physiology

2.1 Morphogenesis and morphology of HF

2.2 Physiology of HF focusing on the hair cycle

3. Experimental approaches to HF regeneration

3.1 Principles of experimental HF regeneration

3.2 Use of hiPSCs for human HF regeneration

4. Relationship between HF and iPSCs: the HF as a favorable iPSC material

5. Regeneration of human HFs using hiPSCs

5.1 Basic principles

5.2 Induction of folliculogenic KCs from hiPSCs

5.3 Generation of dermal cells with trichogenic activity from hiPSCs

5.4 Assembly of HF structures using hiPSC-derived HF components

5.5 Use of hiPSCs for HF formation mimicking normal organogenesis

6. Strategies for treating hair loss disorders using hiPSCs

6.1 Development of pathophysiologically specific therapeutic approaches

6.2 Cell-based therapies to reverse HF miniaturization

6.3 Suppression of immune responses involving HFs by hiPSC-derived cells

7. Conclusion

List of abbreviations

References

1. Introduction

The hair follicle (HF) is a skin appendage and a characteristic of mammals (Montagna and Parakkal, 1974). The HF is a multifunctional miniorgan that provides a physical barrier, acts as sensory machinery, and enables thermoregulation (Paus and Cotsarelis, 1999; Schneider et al., 2009). HFs can influence social interactions (Paus and Cotsarelis, 1999). HFs greatly impact physical appearance in humans (Rose, 2018; Ahluwalia and Fabi, 2019). Accordingly, there is much demand for treatment of hair loss disorders (Rose, 2018; Endo et al., 2018). The so-called androgenetic alopecia is a common form of hair loss and is subdivided into the androgen-dependent male pattern hair loss (MPHL) and less androgen-dependent and more heterogeneous female pattern hair loss (FPHL) (Olsen et al., 2005; Carmina et al., 2019) (Fig. 1.1). Effective oral medications (for MPHL) and autologous HF transplantation (for MPHL and FPHL) are readily available; however, their efficacy is limited for advanced cases (Jung et al., 2014; Adil and Godwin, 2017; Manabe et al., 2018). Alopecia areata (AA) is a commonly encountered autoimmune-mediated hair loss disease (Strazzulla et al., 2018a) (Fig. 1.2). Despite the development of potentially groundbreaking drugs for severe cases, no treatment has been approved by the Food and Drug Administration (Strazzulla et al., 2018b; Jabbari et al., 2018). Furthermore, the treatment options for other forms of intractable alopecia, such as congenital hypotrichosis, primary scarring alopecia leaving permanent hair loss, and advanced-stage FPHL, are limited (Manabe et al., 2018; Kinoshita-Ise et al., 2017; Harries et al., 2008).

Figure 1.1 Clinical and trichoscopic findings of MPHL and FPHL.MPHL and FPHL are characterized by the distribution of hair loss areas (the frontotemporal angle and the vertex in MPHL and the crown and the frontal area with retention of the frontal hairline in FPHL); however, both conditions share the same pathology of hair miniaturization as detected by trichoscopic examination.

Figure 1.2 Clinicopathological findings of AA.(a) Typically, AA is characterized by round to oval patches of hair loss, which can merge to affect larger areas. Total scalp or body hair loss can be observed. (b) AA is an autoimmune disease. Peribulbar lymphocytic infiltration (arrows) is a histopathological hallmark of AA.

Methods for experimental HF regeneration have been developed (Ohyama et al., 2010; Yang and Cotsarelis, 2010; Ohyama and Veraitch, 2013; Ohyama, 2019). Stem cell/progenitor populations of HF components, which maintain continuous HF self-renewal cycles, have been identified and isolated as living cells (Ohyama and Veraitch, 2013; Ohyama, 2019). Use of such plastic and highly proliferative cells should facilitate full establishment of human HF reproduction technology, which is hampered by major technical hurdles. Among them, the shortage of starting human HF-derived materials and the loss of their HF-prone intrinsic properties during in vitro expansion are important (Ohyama and Veraitch, 2013; Ohyama, 2019).

Human induced pluripotent stem cells (human iPSCs or hiPSCs), with their theoretically unlimited proliferative nature and the capacity to differentiate into multiple cell lineages, may provide strategies to overcome the obstacles to human HF regeneration (Ohyama and Veraitch, 2013; Ohyama, 2019). In this chapter, HF morphology and physiology and previously established experimental HF regeneration techniques are first introduced. Next, past and current investigations using hiPSCs for HF regeneration are described, with a focus on the advantage of hiPSCs over conventionally adopted HF-derived cell subsets. Finally, the pathophysiology of hair loss disorders and possible hiPSC-based remedies for individual conditions, including alternatives to HF regeneration, are explained.

2. HF morphology and physiology

2.1. Morphogenesis and morphology of HF

HF morphogenesis depends on well-orchestrated epithelial–mesenchymal interactions (EMIs), which begin with the formation of the placode, a focal thickening of the fetal epithelium, which is driven by WNT signals arising from the underlying dermis (Sennett and Rendl, 2012; Saxena et al., 2019) (Fig. 1.3). In response to the dermal signals from a cell condensate (the dermal condensate) formed beneath the placode, the fetal epithelium and the basement membrane invaginate into the dermis to form a hollow cylindrical structure consisting of multiple layers of keratinocytes (KCs). This structure is surrounded by a collagenous connective tissue sheath (the dermal sheath; DS), to form the main body of the HF (Figs. 1.3 and 1.4) (Paus and Cotsarelis, 1999; Sennett and Rendl, 2012; Stenn and Paus, 2001; Muller-Rover et al., 2001). The epithelial main body is divided into two major layers; the outer root sheath (ORS) and the inner root sheath, which are separated by the biochemically distinct companion layer (Fig. 1.4) (Stenn and Paus, 2001; Muller-Rover et al., 2001; Sperling, 1991). A highly specialized mesenchymal cell aggregate, the dermal papilla (DP), is located at the root of the HF bilaterally surrounded by ORS and capped by hair matrix cells, a highly proliferative KC subset, which divide in response to DP signals to extend the hair shaft (Paus and Cotsarelis, 1999; Stenn and Paus, 2001) (Fig. 1.4). Melanocytes reside around the DP–hair matrix border (Sperling, 1991). The HF has several subsidiary structures, such as the sebaceous gland and the arrector pili muscle (Stenn and Paus, 2001; Muller-Rover et al., 2001; Sperling, 1991) (Fig. 1.4). The insertion point of the arrector pili muscle of ORS is termed the bulge and harbors HF epithelial stem cells (Cotsarelis et al., 1990; Lyle et al., 1998; Ohyama et al., 2006).

Figure 1.3 HF morphogenesis and the hair cycle.HF morphogenesis are enabled by well-orchestrated epithelial–mesenchymal interactions (EMI; arrows). HFs self-renew via the hair cycle consisting of anagen (growing phase), catagen (regressing phase), telogen (resting phase), and exogen (hair-shedding phase) stages, regulated by DP.

Figure 1.4 Morphological and histological characteristics of the HF.Microdissected human anagen HF (left panel) and histopathological images (middle panel; low magnification, right panel; close-up image of the bulb portion).

2.2. Physiology of HF focusing on the hair cycle

Each adult HF periodically and randomly regenerates itself via the hair cycle. This consists of the anagen (growing phase), catagen (regression phase), telogen (resting phase), and exogen (hair-shedding phase) stages (Fig. 1.3) (Paus and Cotsarelis, 1999; Stenn and Paus, 2001; Muller-Rover et al., 2001). The aforementioned structural characteristics (Fig. 1.4) are those of anagen HFs, which predominate in the scalp and therefore are clinically important. In the case of human HFs, after several years of anagen phase, the proximal portion of the HF below the bulge regresses by apoptosis during the 2–3 weeks of the catagen stage, leaving the distal HF portion above the bulge (Fig. 1.3) (Paus and Cotsarelis, 1999; Stenn and Paus, 2001; Kligman, 1959). In the following telogen stage, which lasts for around 2 months, the proximal end of the hair shaft is fully keratinized, allowing hair shedding in the subsequent exogen stage (Stenn and Paus, 2001; Higgins et al., 2009). During this process, DP relocates near the bulge, presumably enabling DP cells (DPCs) to communicate with bulge epithelial stem cells to initiate the next round of the hair cycle by inducing the anagen stage (Panteleyev et al., 2001).

The hair cycle continues throughout life and is maintained by stem/progenitor cell populations of key HF components, such as bulge epithelial stem cells, subbulge melanocyte stem cells, and DPC precursors residing in the cup-shaped bottom portion of the DS (DS cup cells; DSCCs) (Fig. 1.4) (Ohyama and Veraitch, 2013; Ohyama, 2019; Rahmani et al., 2014; Nishimura et al., 2002).

Dysregulation of the hair cycle, e.g., anagen HFs’ sudden and simultaneous entry into telogen, results in a pathological increase in hair shedding (telogen effluvium) (Harrison and Sinclair, 2002). In addition, acceleration of the hair cycle accompanied by shortening of the anagen stage leads to HF miniaturization, which is typical of MPHL and FPHL (Fig. 1.1) (Manabe et al., 2018; Whiting, 2001; Messenger and Sinclair, 2006).

3. Experimental approaches to HF regeneration

3.1. Principles of experimental HF regeneration

Various approaches to experimental HF regeneration have been reported (Ohyama et al., 2010; Yang and Cotsarelis, 2010; Ohyama and Veraitch, 2013; Ohyama, 2019). Most methodologies involve modifications of the same principle—cografting of epithelial and trichogenic dermal (mesenchymal) cells into in vivo permissive environments, e.g., the subcutaneous spaces or under the kidney capsule, of immunodeficient mice (Fig. 1.5).

The chamber (epithelial and dermal cells are mixed and injected into silicone chambers transplanted onto the back of a mouse) (Lichti et al., 1993; Weinberg et al., 1993) and patch (a small amount of cell mixture is grafted into the subcutaneous space) (Zheng et al., 2005) techniques are the standard methods (Fig. 1.5). Because HF formation requires EMIs (Fig. 1.3) (Sennett and Rendl, 2012; Saxena et al., 2019), the absence of the epithelial or dermal component would prevent HF regeneration (Ohyama and Veraitch, 2013; Ohyama, 2019). These assays usually achieve a satisfactory HF yield using murine cells (Weinberg et al., 1993; Zheng et al., 2005). Of note, use of bulge KCs or freshly isolated trichogenic dermal cells improves the regeneration rate (Morris et al., 2004; Blanpain et al., 2004; Ito et al., 2007), suggesting that both the receptivity of epithelial cells to hair-inductive dermal signals and the intensity of trichogenic activity in dermal cells are pivotal determinants of the magnitude of folliculogenic EMIs (Ohyama and Veraitch, 2013; Ohyama, 2019).

Figure 1.5 Major approaches for HF regeneration.The chamber assay and the path assay are the gold standard methods for HF regeneration. In both assays, epithelial and mesenchymal cells are mixed and cografted in vivo, typically into immunodeficient mice. HFs are formed by spontaneous cell–cell reassembly (arrowheads).

3.2. Use of hiPSCs for human HF regeneration

Unlike murine HFs, experimental regeneration of human HFs is technically difficult. Indeed, adaptation of the aforementioned experimental approaches to human HF regeneration is often limited by the paucity of starting materials and the loss of cell properties optimal for HF induction during in vitro expansion (Ohyama and Veraitch, 2013; Ohyama, 2019). hiPSCs, with their theoretically unlimited proliferative properties and ability to differentiate into multiple cell lineages, show promise for overcoming these technical hurdles (Ohyama, 2019; Ohyama and Okano, 2013).

4. Relationship between HF and iPSCs: the HF as a favorable iPSC material

HFs are easily accessible and can be collected less invasively. Multiple cellular components of HFs, e.g., ORS KCs, melanocytes, and DPCs, are reported to be reprogrammable as hiPSCs (Ohyama, 2019). Because these components can be collected from a successfully plucked HF (Petit et al., 2012) and the differentiation potential of hiPSC lines varies according to their origins, the HF may be a favorable material for hiPSC generation, especially for HF regeneration purposes (Ohyama, 2019).

Intrinsic upregulation of several Yamanaka factors (Oct3/4, Sox2, Klf4, c-Myc) was observed in melanocytes (Sox2) and in murine DPCs (Sox2 and Klf4) (Utikal et al., 2009; Tsai et al., 2011). Taking advantage of this preferential gene expression profile, murine DPCs were reprogrammed into iPSCs by introduction of Oct4 alone (Tsai et al., 2011). Although human DPCs require transduction of all four Yamanaka factors to generate hiPSCs, the efficiency of DPC reprogramming was up to 0.03%, while that of dermal fibroblasts under the same condition was up to 0.01%, supporting the suitability of DPCs for hiPSC generation (Higgins et al., 2012; Muchkaeva et al., 2014). Also, an improved protocol for reprogramming plucked HF-derived KCs (HFKCs) into hiPSC without feeder cells and serum-containing medium using integration-free Sendai virus has been established (Re et al., 2018), highlighting the advantage of the HF as a starting material for hiPSC generation.

5. Regeneration of human HFs using hiPSCs

5.1. Basic principles

A method of regenerating HFs from hiPSCs would theoretically allow infinite HF self-replication by reprogramming HF cells into hiPSCs (Ohyama, 2019). Expansion of HF-derived hiPSCs and redifferentiation into HF cell subsets may enable highly efficient regenerative treatments for hair loss disorders. To that end, two approaches can be conceived (Fig. 1.6): (1) preparation of individual HF cell components followed by reconstruction of the HF structure using prepared materials (Ohyama, 2019) and (2) direct HF regeneration from hiPSCs mimicking normal organogenesis (Takagi et al., 2016). The former would facilitate both optimization of each component to enhance the HF regeneration efficiency and adjustment of the size of the regenerated structures. Additional applications, e.g., hiPSC-derived HF-KC and DPC coculture for drug discovery, would also be possible using this approach. However, this approach has multiple steps and is technically challenging (Ohyama, 2019). The latter approach would be more physiological, straightforward, and stable. However, modification of the hiPSC-derived product during the regeneration process, e.g., size or shape adjustment, can be more technically difficult than using the former approach (Ohyama, 2019). We favor the first strategy and have attempted to generate each HF component by a stepwise approach (Ohyama, 2019; Veraitch et al., 2013, 2017) (Fig. 1.6).

Figure 1.6 Approaches for experimental regeneration of human HFs using hiPSCs.HF epithelial and hair-inductive dermal cell components are induced from hiPSCs to construct the main body of HF and DPC aggregates, respectively, which assemble to regenerate HF structures. Alternatively, hiPSCs-derived embryoid bodies are implanted in vivo (e.g., into immunodeficient mice) to recapitulate organogenesis to give rise to organoids containing HFs, which are isolated for downstream applications. 3D-IOS, 3D integumentary organ system; HF, hair follicles. Three-dimensional images were obtained in a pilot study conducted under JSPS KAKENHI: JP 16H05370.

5.2. Induction of folliculogenic KCs from hiPSCs

Based on the protocol for KC generation from human embryonic stem cells using retinoic acid (RA) and BMP4 (Metallo et al., 2008), well-differentiated KCs have been induced from hiPSCs (Fig. 1.6a, 7a ) (Itoh et al., 2011, 2013). Neonatal or fetal human KCs were more receptive to hair-inductive dermal signals than adult KCs, which formed HF structures in in vivo reconstitution assays (Ehama et al., 2007; Thangapazham et al., 2014). These observations imply that use of juvenile and putatively plastic KCs enhances the stability of human HF regeneration.

Because most hair loss patients are adults, providing aged KCs, attempts have been made to endow KCs with HF characteristics by exposure to activators of the signaling pathways essential to HF development (e.g., WNT, SHH, ectodysplasin-A, and BMP (Saxena et al., 2019)) or epidermal and basic fibroblast growth factors. These efforts have met with some success (Sun et al., 2011).

Because hiPSCs recapitulate fetal KC differentiation during their differentiation into KCs, progenitors with biological properties analogous to those of fetal KCs with high receptivity to trichogenic dermal signals can be generated (Ohyama and Veraitch, 2013; Veraitch et al., 2013).

In line with this hypothesis, ectodermal precursor cells (EPCs), which were generated by exposing hiPSCs to RA-BMP4 and collected without passaging in KC culture medium, expressed both keratin 14 and 18 (Fig. 1.7b), suggesting them as being innate (Veraitch et al., 2013). When cocultured with human DPCs, one EPC line more strongly upregulated HF-related keratin genes than normal human adult interfollicular KCs (Veraitch et al., 2013). That EPC line also showed enhanced