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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.

Zhao Y, Jiang Z, Zhao T, Ye M, Hu C, Zhou H, Yin Z, Chen Y, Zhang Y, Wang S, Shen J, Thaker H, Jain S, Li Y, Diao Y, Chen Y, Sun X, Fisk MB, Li H - BMC Med (2013)

Bottom Line: Clinical findings indicate that T2D patients achieve improved metabolic control and reduced inflammation markers after receiving Stem Cell Educator therapy.Median glycated hemoglobin (HbA1C) in Group A and B was significantly reduced from 8.61%±1.12 at baseline to 7.25%±0.58 at 12 weeks (P=2.62E-06), and 7.33%±1.02 at one year post-treatment (P=0.0002).In addition, this approach does not appear to have the safety and ethical concerns associated with conventional stem cell-based approaches.

View Article: PubMed Central - HTML - PubMed

Affiliation: Section of Endocrinology, Diabetes and Metabolism, Department of Medicine, University of Illinois at Chicago, 1819 W, Polk Street, Chicago, IL 60612, USA. yzhaowhl@yahoo.com

ABSTRACT

Background: 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.

Methods: In an open-label, phase 1/phase 2 study, patients (N=36) with long-standing T2D were divided into three groups (Group A, oral medications, n=18; Group B, oral medications+insulin injections, n=11; Group C having impaired β-cell function with oral medications+insulin injections, n=7). All patients received one treatment with the Stem Cell Educator therapy in which a patient's blood is circulated through a closed-loop system that separates mononuclear cells from the whole blood, briefly co-cultures them with adherent cord blood-derived multipotent stem cells (CB-SCs), and returns the educated autologous cells to the patient's circulation.

Results: Clinical findings indicate that T2D patients achieve improved metabolic control and reduced inflammation markers after receiving Stem Cell Educator therapy. Median glycated hemoglobin (HbA1C) in Group A and B was significantly reduced from 8.61%±1.12 at baseline to 7.25%±0.58 at 12 weeks (P=2.62E-06), and 7.33%±1.02 at one year post-treatment (P=0.0002). Homeostasis model assessment (HOMA) of insulin resistance (HOMA-IR) demonstrated that insulin sensitivity was improved post-treatment. Notably, the islet beta-cell function in Group C subjects was markedly recovered, as demonstrated by the restoration of C-peptide levels. Mechanistic studies revealed that Stem Cell Educator therapy reverses immune dysfunctions through immune modulation on monocytes and balancing Th1/Th2/Th3 cytokine production.

Conclusions: Clinical data from the current phase 1/phase 2 study demonstrate that Stem Cell Educator therapy is a safe approach that produces lasting improvement in metabolic control for individuals with moderate or severe T2D who receive a single treatment. In addition, this approach does not appear to have the safety and ethical concerns associated with conventional stem cell-based approaches.

Trial registration: ClinicalTrials.gov number, NCT01415726.

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Related in: MedlinePlus

In vitro study of the immune modulation of CB-SCs on monocytes. (A) Phase contrast microscopy shows the co-culture of CB-SC with monocytes (bottom left panel) for 18 hrs. CB-SCs co-culture with lymphocytes (top right panel) served as the control. The impaired CB-SCs after co-culture with monocytes were restored to expansion and became 90 to approximately 100% confluence after 7 to 10 days (bottom right). Original magnification, × 100. (B) Apoptotic analysis of floating cells from the co-culture of CB-SCs with monocytes for 18 hrs. (C) Western blotting shows the expression of the cellular inhibitor of apoptosis protein (cIAP) 1, not cIAP2, in four preparations of CB-SCs. (D) Western blotting shows the expression of tumor necrosis factor receptor II (TNF-RII), not TNF-RI, in four preparations of CB-SCs. (E) TNFα suppresses the proliferation of CB-SCs in a dose–response manner. Cell proliferation was evaluated using CyQUANTR Cell Proliferation Assay Kit [25]. (F) The blocking experiment with iNOS inhibitor 1400W demonstrates that CB-SC-derived nitric oxide (NO) contributes to the immune modulation of CB-SCs on monocytes. Monocytes were initially stimulated with lipopolysaccharide (LPS, 10 μg/ml) for 8 hrs, and then co-cultured with CB-SCs at ratio 1:5 of CB-SCs:monocytes for 48 hrs in the presence or absence of 1400W (100 nM), followed by real time PCR analysis by using Human Th17 for Autoimmunity and Inflammation PCR Array kit (SABiosciences, Valencia, CA, USA).
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Figure 3: In vitro study of the immune modulation of CB-SCs on monocytes. (A) Phase contrast microscopy shows the co-culture of CB-SC with monocytes (bottom left panel) for 18 hrs. CB-SCs co-culture with lymphocytes (top right panel) served as the control. The impaired CB-SCs after co-culture with monocytes were restored to expansion and became 90 to approximately 100% confluence after 7 to 10 days (bottom right). Original magnification, × 100. (B) Apoptotic analysis of floating cells from the co-culture of CB-SCs with monocytes for 18 hrs. (C) Western blotting shows the expression of the cellular inhibitor of apoptosis protein (cIAP) 1, not cIAP2, in four preparations of CB-SCs. (D) Western blotting shows the expression of tumor necrosis factor receptor II (TNF-RII), not TNF-RI, in four preparations of CB-SCs. (E) TNFα suppresses the proliferation of CB-SCs in a dose–response manner. Cell proliferation was evaluated using CyQUANTR Cell Proliferation Assay Kit [25]. (F) The blocking experiment with iNOS inhibitor 1400W demonstrates that CB-SC-derived nitric oxide (NO) contributes to the immune modulation of CB-SCs on monocytes. Monocytes were initially stimulated with lipopolysaccharide (LPS, 10 μg/ml) for 8 hrs, and then co-cultured with CB-SCs at ratio 1:5 of CB-SCs:monocytes for 48 hrs in the presence or absence of 1400W (100 nM), followed by real time PCR analysis by using Human Th17 for Autoimmunity and Inflammation PCR Array kit (SABiosciences, Valencia, CA, USA).

Mentions: To better understand the immune modulation of CB-SC on monocytes, we performed in vitro co-culture experiments by using CD14+ monocytes purified from human peripheral blood. The purified CD14+ monocytes were co-cultured with CB-SCs at different ratios. We found that there were strong reactions after adding the CD14+ monocytes to CB-SCs (Figure 3A, bottom left panel). Flow analysis demonstrated that co-culture with CB-SCs for 18 hrs resulted in the significant apoptosis of monocytes at the ratio 1:5 of CB-SC:monocytes (Figure 3B). Correspondingly, both the cell viability and attachment of CB-SCs were also affected in the presence of apoptotic monocytes (Figure 3A, bottom left panel). The cellular processes of CB-SCs were reduced in length, but most were still attached to the bottom (Figure 3A, bottom left panel). Interestingly, these impaired CB-SCs were restored after co-culture for 2 to 3 days; they continually expanded and became 90 to approximately 100% confluence after 7 to 10 days (Figure 3A, bottom right panel). Mechanistic studies revealed that CB-SCs displayed the cellular inhibitor of apoptosis protein (cIAP) 1 [41] that protects CB-SCs against the cytotoxic effects of monocytes, allowing them to survive and proliferate (Figure 3C). To further explore the molecular mechanisms underlying the cytotoxic effects of monocytes on CB-SCs, we found that CB-SCs expressed TNF-RII but not TNF-RI (Figure 3D). Recombinant TNF showed cytotoxicity to CB-SCs at different doses (Figure 3E). Notably, CB-SCs pre-treated with TNF-RII mAb (20 μg/ml) at a ratio of 1:10 could markedly block the toxic action of monocytes and protect 50% of CB-SCs with good cell viability and morphology.


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.

Zhao Y, Jiang Z, Zhao T, Ye M, Hu C, Zhou H, Yin Z, Chen Y, Zhang Y, Wang S, Shen J, Thaker H, Jain S, Li Y, Diao Y, Chen Y, Sun X, Fisk MB, Li H - BMC Med (2013)

In vitro study of the immune modulation of CB-SCs on monocytes. (A) Phase contrast microscopy shows the co-culture of CB-SC with monocytes (bottom left panel) for 18 hrs. CB-SCs co-culture with lymphocytes (top right panel) served as the control. The impaired CB-SCs after co-culture with monocytes were restored to expansion and became 90 to approximately 100% confluence after 7 to 10 days (bottom right). Original magnification, × 100. (B) Apoptotic analysis of floating cells from the co-culture of CB-SCs with monocytes for 18 hrs. (C) Western blotting shows the expression of the cellular inhibitor of apoptosis protein (cIAP) 1, not cIAP2, in four preparations of CB-SCs. (D) Western blotting shows the expression of tumor necrosis factor receptor II (TNF-RII), not TNF-RI, in four preparations of CB-SCs. (E) TNFα suppresses the proliferation of CB-SCs in a dose–response manner. Cell proliferation was evaluated using CyQUANTR Cell Proliferation Assay Kit [25]. (F) The blocking experiment with iNOS inhibitor 1400W demonstrates that CB-SC-derived nitric oxide (NO) contributes to the immune modulation of CB-SCs on monocytes. Monocytes were initially stimulated with lipopolysaccharide (LPS, 10 μg/ml) for 8 hrs, and then co-cultured with CB-SCs at ratio 1:5 of CB-SCs:monocytes for 48 hrs in the presence or absence of 1400W (100 nM), followed by real time PCR analysis by using Human Th17 for Autoimmunity and Inflammation PCR Array kit (SABiosciences, Valencia, CA, USA).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC3716981&req=5

Figure 3: In vitro study of the immune modulation of CB-SCs on monocytes. (A) Phase contrast microscopy shows the co-culture of CB-SC with monocytes (bottom left panel) for 18 hrs. CB-SCs co-culture with lymphocytes (top right panel) served as the control. The impaired CB-SCs after co-culture with monocytes were restored to expansion and became 90 to approximately 100% confluence after 7 to 10 days (bottom right). Original magnification, × 100. (B) Apoptotic analysis of floating cells from the co-culture of CB-SCs with monocytes for 18 hrs. (C) Western blotting shows the expression of the cellular inhibitor of apoptosis protein (cIAP) 1, not cIAP2, in four preparations of CB-SCs. (D) Western blotting shows the expression of tumor necrosis factor receptor II (TNF-RII), not TNF-RI, in four preparations of CB-SCs. (E) TNFα suppresses the proliferation of CB-SCs in a dose–response manner. Cell proliferation was evaluated using CyQUANTR Cell Proliferation Assay Kit [25]. (F) The blocking experiment with iNOS inhibitor 1400W demonstrates that CB-SC-derived nitric oxide (NO) contributes to the immune modulation of CB-SCs on monocytes. Monocytes were initially stimulated with lipopolysaccharide (LPS, 10 μg/ml) for 8 hrs, and then co-cultured with CB-SCs at ratio 1:5 of CB-SCs:monocytes for 48 hrs in the presence or absence of 1400W (100 nM), followed by real time PCR analysis by using Human Th17 for Autoimmunity and Inflammation PCR Array kit (SABiosciences, Valencia, CA, USA).
Mentions: To better understand the immune modulation of CB-SC on monocytes, we performed in vitro co-culture experiments by using CD14+ monocytes purified from human peripheral blood. The purified CD14+ monocytes were co-cultured with CB-SCs at different ratios. We found that there were strong reactions after adding the CD14+ monocytes to CB-SCs (Figure 3A, bottom left panel). Flow analysis demonstrated that co-culture with CB-SCs for 18 hrs resulted in the significant apoptosis of monocytes at the ratio 1:5 of CB-SC:monocytes (Figure 3B). Correspondingly, both the cell viability and attachment of CB-SCs were also affected in the presence of apoptotic monocytes (Figure 3A, bottom left panel). The cellular processes of CB-SCs were reduced in length, but most were still attached to the bottom (Figure 3A, bottom left panel). Interestingly, these impaired CB-SCs were restored after co-culture for 2 to 3 days; they continually expanded and became 90 to approximately 100% confluence after 7 to 10 days (Figure 3A, bottom right panel). Mechanistic studies revealed that CB-SCs displayed the cellular inhibitor of apoptosis protein (cIAP) 1 [41] that protects CB-SCs against the cytotoxic effects of monocytes, allowing them to survive and proliferate (Figure 3C). To further explore the molecular mechanisms underlying the cytotoxic effects of monocytes on CB-SCs, we found that CB-SCs expressed TNF-RII but not TNF-RI (Figure 3D). Recombinant TNF showed cytotoxicity to CB-SCs at different doses (Figure 3E). Notably, CB-SCs pre-treated with TNF-RII mAb (20 μg/ml) at a ratio of 1:10 could markedly block the toxic action of monocytes and protect 50% of CB-SCs with good cell viability and morphology.

Bottom Line: Clinical findings indicate that T2D patients achieve improved metabolic control and reduced inflammation markers after receiving Stem Cell Educator therapy.Median glycated hemoglobin (HbA1C) in Group A and B was significantly reduced from 8.61%±1.12 at baseline to 7.25%±0.58 at 12 weeks (P=2.62E-06), and 7.33%±1.02 at one year post-treatment (P=0.0002).In addition, this approach does not appear to have the safety and ethical concerns associated with conventional stem cell-based approaches.

View Article: PubMed Central - HTML - PubMed

Affiliation: Section of Endocrinology, Diabetes and Metabolism, Department of Medicine, University of Illinois at Chicago, 1819 W, Polk Street, Chicago, IL 60612, USA. yzhaowhl@yahoo.com

ABSTRACT

Background: 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.

Methods: In an open-label, phase 1/phase 2 study, patients (N=36) with long-standing T2D were divided into three groups (Group A, oral medications, n=18; Group B, oral medications+insulin injections, n=11; Group C having impaired β-cell function with oral medications+insulin injections, n=7). All patients received one treatment with the Stem Cell Educator therapy in which a patient's blood is circulated through a closed-loop system that separates mononuclear cells from the whole blood, briefly co-cultures them with adherent cord blood-derived multipotent stem cells (CB-SCs), and returns the educated autologous cells to the patient's circulation.

Results: Clinical findings indicate that T2D patients achieve improved metabolic control and reduced inflammation markers after receiving Stem Cell Educator therapy. Median glycated hemoglobin (HbA1C) in Group A and B was significantly reduced from 8.61%±1.12 at baseline to 7.25%±0.58 at 12 weeks (P=2.62E-06), and 7.33%±1.02 at one year post-treatment (P=0.0002). Homeostasis model assessment (HOMA) of insulin resistance (HOMA-IR) demonstrated that insulin sensitivity was improved post-treatment. Notably, the islet beta-cell function in Group C subjects was markedly recovered, as demonstrated by the restoration of C-peptide levels. Mechanistic studies revealed that Stem Cell Educator therapy reverses immune dysfunctions through immune modulation on monocytes and balancing Th1/Th2/Th3 cytokine production.

Conclusions: Clinical data from the current phase 1/phase 2 study demonstrate that Stem Cell Educator therapy is a safe approach that produces lasting improvement in metabolic control for individuals with moderate or severe T2D who receive a single treatment. In addition, this approach does not appear to have the safety and ethical concerns associated with conventional stem cell-based approaches.

Trial registration: ClinicalTrials.gov number, NCT01415726.

Show MeSH
Related in: MedlinePlus