Research in the Zhao Lab
Stem Cell Educator therapy in type 1 diabetes (T1D)
T1D is a T cell-mediated autoimmune disease that causes the deficit of pancreatic islet b cells. A true cure has proven elusive despite intensive research using conventional approaches over the past 25 years. The situation highlights the challenges we face in conquering this disease. Alternative approaches are needed. Increasing evidence demonstrates that stem cells possess the function of immune modulation. We developed Stem Cell Educator therapy using cord blood-derived multipotent stem cells (CB-SCs). A closed-loop system that circulates a patient’s blood through a blood cell separator briefly co-cultures the patient’s lymphocytes with adherent CB-SCs in vitro, and returns the educated lymphocytes (but not the CB-SCs) to the patient’s circulation. Our clinical trial revealed that a single treatment with Stem Cell Educator therapy provides lasting reversal of autoimmunity that allows regeneration of islet b cells and improvement of metabolic control in patients with longstanding T1D.
Overview of Stem Cell Educator Therapy and Related Mechanisms. A participant (left) is connected to a Blood Cell Separator (right) and the Stem Cell Educator (bottom center) to form a closed system. Lymphocytes isolated from the T1D participant by the Blood Cell Separator travel through the Stem Cell Educator, where they come in contact with CB-SCs attached to the interior surfaces of the device, through the autoimmune regulator (AIRE) and other related mechanisms modulating immune cells. Educated lymphocytes are returned to the patient’s blood circulation.
Improvement of β-cell function by Stem Cell Educator therapy. (A) Fasting C-peptide levels of T1D participants over 24 weeks. Group A (with some residual β-cell function) and Group B (severe longstanding subjects with no or some residual β-cell function) participants (n = 6 per group) received one Stem Cell Educator treatment. Control group participants (n = 3) received sham therapy (no CB-SCs in the Stem Cell Educator). (B) Comparison of C-peptide levels at glucose challenge after 40-week follow-up in Group B T1D subjects. The dashed red line indicates the lower limit for normal C-peptide levels in Chinese populations. The dashed purple line indicates the minimum detectable level (sensitivity) of C-peptide by radioimmunoassay (RIA). Abbreviations: CB-SCs, cord blood stem cells; OGTT, oral glucose tolerance test; T1D, type 1 diabetes.
Delgado E, Perez-Basterrechea M, Suarez-Alvarez B, Zhou H, Martinez Revuelta E, Garcia-Gala JM, Perez S, Avrez-Viejo M, Menendez E, Lopez-Larrea C, Tang R, Zhu Z, Hu W, Moss T, Guindi E, Otero J, and Zhao Y. Modulation of autoimmune T-Cell memory by Stem Cell Educator therapy: phase 1/2 clinical trial. EBioMedicine 2015, 2, 2024-2036.
Yong Zhao. Stem Cell Educator Therapy and Induction of Immune Balance. Current Diabetes Reports. 2012, Jul.26, [Epub ahead of print]; 12(5):517-23.
Yong Zhao, Zhaoshun Jiang, Tingbao Zhao, Mingliang Ye, Chengjin Hu, Zhaohui Yin, Heng Li, Ye Zhang, Yalin Diao, Yunxiang Li, Yingjian Chen, Xiaoming Sun, Mary Beth Fisk, Randal A. Skidgel, Mark Holterman, Bellur Prabhakar, Theodore Mazzone. Reversal of Type 1 Diabetes via Islet beta Cell Regeneration Following Immune Modulation by Cord Blood-Derived Multipotent Stem Cells. BMC Medicine. 2012 Jan 10;10(1): 3.
Yong Zhao, Theodore Mazzone. Human cord blood stem cells and the journey to a cure for type 1 diabetes. Autoimmunity Reviews. 10:103-107, 2010.
Yong Zhao, Brian Lin, Robert Darflinger, Yongkang Zhang, Mark Holterman, Randal A. Skidgel. Human cord blood stem cell-modulated regulatory T lymphocytes reverse the autoimmune-caused type 1 diabetes in nonobese diabetic (NOD) mice. Plos ONE. 4 (1): e4226, 2009.
Xiang Song, Wei Hu, Haibo Yu, Honglan Wang, Yelu Zhao, Robert Korngold, Yong Zhao*. Existence of circulating mitochondria in human and animal peripheral blood. International Journal of Molecular Science 2020; 21, 2122. PMID: 32204530
Wei Hu, Xiang Song, Haibo Yu, Jingyu Sun, Yong Zhao*. Released Exosomes Contribute to the Immune Modulation of Cord Blood-Derived Stem Cells (CB-SC). Frontiers in Immunology. 2020; 11: 165. PMID: 32161585
Stem Cell Educator therapy in type 2 diabetes (T2D)
The prevalence of type 2 diabetes (T2D) is increasing worldwide and creating a significant burden on health systems, highlighting the need for the development of innovative therapeutic approaches to overcome immune dysfunction, which is likely a key factor in the development of insulin resistance in T2D. It suggests that immune modulation may be a useful tool in treating the disease.
Specifically, inflammatory cytokines derived from adipocytes and macrophages promote the development of insulin resistance in T2D through JNK and/or IKK/NF-B pathways, including changes in the levels of tumor necrosis factor- (TNF), interleukin-1 (IL-1), IL-6, IL-17, monocyte chemoattractant protein-1 (MCP-1), resistin, and plasminogen activator inhibitor-1 (PAI-1). Control or reversal of these immune dysfunctions and chronic inflammation may provide an alternative approach for overcoming insulin resistance and may point to a cure for diabetes. The failure of several recent clinical trials in type 1 diabetes (T1D), however, highlights the challenges we face in conquering multiple immune dysfunctions by using conventional immune approaches in patients.
Stem Cell Educator therapy is an innovative technology we developed and is designed to control or reverse immune dysfunction. We explored the therapeutic potential of Stem Cell Educator therapy in patients with T2D. Clinical data from a Phase I/II study demonstrated that Stem Cell Educator therapy is a safe approach that results in lasting improvement in metabolic control for individuals with moderate or severe T2D who received a single treatment.
Improvement of metabolic control by stem cell educator therapy. (A) Analysis of insulin sensitivity by HOMA-IR C-peptide at four weeks post-treatment with Stem Cell Educator therapy. (B) 56-week follow-up C-peptide levels in longstanding T2D subjects with impaired islet β cell function. (C) Comparison of fasting and high glucose-stimulated C-peptide levels after 4-year follow-up in longstanding severe T2D subjects (N = 4). Data are presented as mean ± SD. Abbreviation: OGTT, oral glucose tolerance test.
Yong Zhao, Zhaoshun Jiang, Tingbao Zhao, Mingliang Ye, Chengjin Hu, Huimin Zhou, Zhaohui Yin, Yana Chen, Ye Zhang, Shanfeng Wang, Jie Shen, Hatim Thaker, Summit Jain, Yunxiang Li, Yalin Diao, Yingjian Chen, Xiaoming Sun, Mary Beth Fisk, Heng Li. Targeting insulin resistance in type 2 diabetes via immune modulation of cord blood-derived multipotent stem cells (CB-SCs) in stem cell educator therapy: phase I/II clinical trial. BMC Medicine. 11: 160, 2013.
Yong Zhao, Zhaoshun Jiang, Chengshan Guo. New hope for type 2 diabetics: targeting insulin resistance through the immune modulation of stem cells. Autoimmunity Reviews. 11: 137-142, 2011.
Stem Cell Educator therapy in alopecia areata (AA)
Alopecia areata (AA) is one of the most common autoimmune diseases that targets hair follicles, with high impact on the quality of life and self-esteem of patients due to hair loss. Clinical management and outcomes are challenged by current limited immunosuppressive and immunomodulating regimens. In an open-label Phase I/II study, patients (N = 9) with severe AA received one treatment with Stem Cell Educator therapy. Median age was 20 years (median alopecic duration, 5 years).
Clinical data demonstrated that patients with severe AA achieved improved hair regrowth and quality of life after receiving Stem Cell Educator therapy. Flow cytometry revealed the up-regulation of Th2 cytokines and restoration of balancing Th1/Th2/Th3 cytokine production in the peripheral blood of AA subjects. Immunohistochemistry indicated the formation of a “ring of transforming-growth factor 1 (TGF-1)” around the hair follicles, leading to the restoration of immune privilege of hair follicles and the protection of newly generated hair follicles against autoimmune destruction. Mechanistic studies revealed that co-culture with CB-SC may up-regulate the expression of co-inhibitory molecules BTLA and PD1 on CD8+NKG2D+ effector T cells and suppress their proliferation via HVEM and PD-L1 ligands on CB-SCs.
Clinical data demonstrated the safety and efficacy of Stem Cell Educator therapy for the treatment of AA. This innovative approach produced lasting improvement in hair regrowth in subjects with moderate or severe AA.
Regrowth of hair following Stem Cell Educator therapy. (A) A subject with severe AA achieved complete hair regrowth at 12 week follow-up after receiving Stem Cell Educator therapy and maintained regrowth through the last follow-up (2 years). (B) H&E staining of scalp tissues. (C) Fresh skin tissues were collected from the scalps via biopsy for immunohistochemistry testing in participants with alopecia totalis before treatment and 12 weeks after receiving Stem Cell Educator therapy. TGF-β1 staining surrounds a hair follicle of AA participants after receiving Stem Cell Educator therapy, with vertical section of hair follicle (top panels) and horizontal section of hair follicle. Isotype-matched mouse IgG1 served as a negative control for TGF-β1 immunostaining in a serial hair follicle section. Representative images were obtained from five experiments. Scale bar, 25 μm.
Yanjia Li, Baoyong Yan, Hepeng Wang, Heng Li, Quanhai Li, Dong Zhao, Yana Chen, Ye Zhang, Wenxia Li, Jun Zhang, Shanfeng Wang, Jie Shen, Yunxiang Li, Edward Guindi, Yong Zhao. Hair Regrowth in Alopecia Areata Patients Following Stem Cell Educator Therapy. BMC Medicine. 2015 April 20; 13: 87.
Platelet-derived mitochondria display embryonic stem cell markers and improve pancreatic islet β-cell function in humans
Diabetes is a major global health issue, and the number of individuals with type 1 diabetes (T1D) and type 2 diabetes (T2D) increases annually across multiple populations. Research to develop a cure must overcome multiple immune dysfunctions and the shortage of pancreatic islet β cells, but these challenges have proven intractable despite intensive research efforts over the last several decades. Stem Cell Educator therapy has previously been proven safe and effective in Chinese and Spanish subjects for the improvement of T1D, T2D, and other autoimmune diseases.
Here, four-year follow-up studies demonstrated the long-term safety and clinical efficacy of Stem Cell Educator therapy for the treatment of T1D and T2D. Mechanistic studies found that the nature of platelets was modulated in diabetic subjects after receiving this treatment. Platelets and their released mitochondria display immune tolerance-associated markers that can modulate the proliferation and function of immune cells. Notably, platelets also expressed embryonic stem cell- and pancreatic islet β-cell-associated markers encoded by mitochondrial DNA. Using freshly isolated human pancreatic islets, ex vivo studies established that platelet-releasing mitochondria can migrate to pancreatic islets and be taken up by islet β cells, leading to the proliferation and enhancement of islet β-cell functions.
These findings reveal new mechanisms underlying Stem Cell Educator therapy and open up novel avenues to improve the treatment of diabetes.
Platelet-derived mitochondria improve pancreatic islet β-cell function in humans. (A) Kinetic measurements by flow cytometry demonstrated that human pancreatic islet cells take up MitoTracker Deep Red-labeled mitochondria released from platelets (bottom) in Transwell co-culture. (B) Co-culture of freshly isolated whole human pancreatic islets (top chamber) with platelets labeled with mitochondria (bottom chamber) in Transwells. The pore size of transmembrane is 0.4 μm. (C) Flow cytometry indicates that the percentage of Insulin+Ki67+ islet β cells was increased after co-culture with mitochondria in Transwells. Mitochondria were purified from CB-platelets (n = 4).
Yong Zhao, Zhaoshun Jiang, Elias Delgado, Heng Li, Huimin Zhou, Wei Hu, Marcos Perez-Basterrechea, Anna Janostakova, Qidong Tan, Jing Wang, Mao Mao, Zhaohui Yin, Ye Zhang, Ying Li, Quanhai Li, Jing Zhou, Yunxiang Li, Eva Martinez Revuelta, Jose Maria García-Gala, Honglan Wang, Silvia Perez-López, Maria Alvarez-Viejo, Edelmiro Menendez, Thomas Moss, Edward Guindi, Jesus Otero. Platelet-derived mitochondria display embryonic stem cell markers and improve pancreatic islet β-cell function in humans. STEM CELLS Translational Medicine. July 7, 2017, 6: 1684-1697.
We thank the Hackensack University Medical Center Foundation, Mr. Poddar family, Mr. Ludwig family, the American Diabetes Association (ADA), Juvenile Diabetes Research Foundation (JDRF), Jinan 5150 Program, the Ministry of Science and Technology in China, the National Natural Science Foundation of China, and Tianhe Stem Cell Biotechnologies for supporting our current and past basic and clinical research projects.