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Hepatocyte growth factor enhances engraftment and function of nonhuman primate islets.

Fiaschi-Taesch NM, Berman DM, Sicari BM, Takane KK, Garcia-Ocaña A, Ricordi C, Kenyon NS, Stewart AF - Diabetes (2008)

Bottom Line: Adenoviral delivery of hepatocyte growth factor (HGF) to rodent islets improves islet graft survival and function, markedly reducing the number of islets required to achieve glucose control.Unilateral nephrectomy resulted in an immediate return of glucose to baseline diabetic levels.Interestingly, adenoviral DNA, as well as mouse HGF (mHGF) mRNA derived from the adenovirus, were present for 42 days posttransplantation.

View Article: PubMed Central - PubMed

Affiliation: Division of Endocrinology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA. taeschn@dom.pitt.edu

ABSTRACT

Objective: Adenoviral delivery of hepatocyte growth factor (HGF) to rodent islets improves islet graft survival and function, markedly reducing the number of islets required to achieve glucose control. Here, we asked whether these prior observations in rodent models extend to nonhuman primate (NHP) islets.

Research design and methods: NHP islets were transduced with murine (Ad.mHGF) or human (Ad.hHGF) adenoviral HGF (Ad.HGF) at low multiplicity of infection and studied in vitro. To study the function of Ad.HGF-transduced NHP islets in vivo, a renal subcapsular marginal mass islet transplant model was developed in streptozotocin-induced diabetic NOD-SCID mice.

Results: Baseline glucose values were 454.7 +/- 11.3 mg/dl (n = 7). Transplant of 500 NHP islet equivalents (IE) had only a marginal effect on blood glucose (369.1 +/- 9.7 mg/dl, n = 5). In striking contrast, 500 NHP IE transduced with Ad.mHGF promptly and continuously corrected blood glucose (142.0 +/- 6.2 mg/dl, n = 7) for the 6-week duration of the experiment. Unilateral nephrectomy resulted in an immediate return of glucose to baseline diabetic levels. Interestingly, adenoviral DNA, as well as mouse HGF (mHGF) mRNA derived from the adenovirus, were present for 42 days posttransplantation. Surprisingly, transplant of 500 IE with Ad.hHGF, as compared with Ad.mHGF, resulted in only marginal correction of blood glucose, suggesting that human HGF is less efficient than mHGF in this system.

Conclusions: These studies demonstrate that mHGF markedly improves islet transplant outcomes in the highest preclinical species examined to date. HGF has promise as an agent that can improve islet mass and function in transplant models and likely in other models of types 1 and 2 diabetes.

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Differential effects mouse HGF and human HGF on p42/44 mitogen-activated protein kinase activity in primary murine arterial smooth muscle cells. A: Effects of human HGF (hHGF) and mouse HGF (mHGF) peptides at 25 ng/ml on total ERK and phospho-ERK in primary cultures of murine aortic smooth muscle cells. Note that human HGF fails to activate mitogen-activated protein kinase in mouse arterial smooth muscle cells, whereas mouse HGF readily activates mitogen-activated protein kinase. To insure that the human HGF was active, it was also assayed in rat INS-1 cells: the panel on the far right demonstrates that the identical human HGF preparation that failed to activate mitogen-activated protein kinase in the vascular smooth muscle cells (VSMC) was robustly active in INS-1 cells. B: Densitometric quantification of three experiments as outlined in A. These studies demonstrate that whereas the human HGF peptide is active, it is unable to activate mitogen-activated protein kinase in murine arterial smooth muscle cells, in contrast to mouse HGF, which is active in these same cells. NTx, no treatment.
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f8: Differential effects mouse HGF and human HGF on p42/44 mitogen-activated protein kinase activity in primary murine arterial smooth muscle cells. A: Effects of human HGF (hHGF) and mouse HGF (mHGF) peptides at 25 ng/ml on total ERK and phospho-ERK in primary cultures of murine aortic smooth muscle cells. Note that human HGF fails to activate mitogen-activated protein kinase in mouse arterial smooth muscle cells, whereas mouse HGF readily activates mitogen-activated protein kinase. To insure that the human HGF was active, it was also assayed in rat INS-1 cells: the panel on the far right demonstrates that the identical human HGF preparation that failed to activate mitogen-activated protein kinase in the vascular smooth muscle cells (VSMC) was robustly active in INS-1 cells. B: Densitometric quantification of three experiments as outlined in A. These studies demonstrate that whereas the human HGF peptide is active, it is unable to activate mitogen-activated protein kinase in murine arterial smooth muscle cells, in contrast to mouse HGF, which is active in these same cells. NTx, no treatment.

Mentions: Total p42 and p44 extracellular signal–regulated kinase (ERK), and their phosphorylated counterparts, were examined using extracts of primary cultures of mouse arterial smooth muscle treated with mouse HGF (R&D Systems, Minneapolis, MN) or hHGF (RDI, Concord, MA) at doses of 25 μg/ml as we have described in detail previously (15) and as described in the legend to Fig. 8. Murine primary arterial smooth muscle cultures were prepared and documented to represent arterial smooth muscle cells as described previously (16).


Hepatocyte growth factor enhances engraftment and function of nonhuman primate islets.

Fiaschi-Taesch NM, Berman DM, Sicari BM, Takane KK, Garcia-Ocaña A, Ricordi C, Kenyon NS, Stewart AF - Diabetes (2008)

Differential effects mouse HGF and human HGF on p42/44 mitogen-activated protein kinase activity in primary murine arterial smooth muscle cells. A: Effects of human HGF (hHGF) and mouse HGF (mHGF) peptides at 25 ng/ml on total ERK and phospho-ERK in primary cultures of murine aortic smooth muscle cells. Note that human HGF fails to activate mitogen-activated protein kinase in mouse arterial smooth muscle cells, whereas mouse HGF readily activates mitogen-activated protein kinase. To insure that the human HGF was active, it was also assayed in rat INS-1 cells: the panel on the far right demonstrates that the identical human HGF preparation that failed to activate mitogen-activated protein kinase in the vascular smooth muscle cells (VSMC) was robustly active in INS-1 cells. B: Densitometric quantification of three experiments as outlined in A. These studies demonstrate that whereas the human HGF peptide is active, it is unable to activate mitogen-activated protein kinase in murine arterial smooth muscle cells, in contrast to mouse HGF, which is active in these same cells. NTx, no treatment.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2551685&req=5

f8: Differential effects mouse HGF and human HGF on p42/44 mitogen-activated protein kinase activity in primary murine arterial smooth muscle cells. A: Effects of human HGF (hHGF) and mouse HGF (mHGF) peptides at 25 ng/ml on total ERK and phospho-ERK in primary cultures of murine aortic smooth muscle cells. Note that human HGF fails to activate mitogen-activated protein kinase in mouse arterial smooth muscle cells, whereas mouse HGF readily activates mitogen-activated protein kinase. To insure that the human HGF was active, it was also assayed in rat INS-1 cells: the panel on the far right demonstrates that the identical human HGF preparation that failed to activate mitogen-activated protein kinase in the vascular smooth muscle cells (VSMC) was robustly active in INS-1 cells. B: Densitometric quantification of three experiments as outlined in A. These studies demonstrate that whereas the human HGF peptide is active, it is unable to activate mitogen-activated protein kinase in murine arterial smooth muscle cells, in contrast to mouse HGF, which is active in these same cells. NTx, no treatment.
Mentions: Total p42 and p44 extracellular signal–regulated kinase (ERK), and their phosphorylated counterparts, were examined using extracts of primary cultures of mouse arterial smooth muscle treated with mouse HGF (R&D Systems, Minneapolis, MN) or hHGF (RDI, Concord, MA) at doses of 25 μg/ml as we have described in detail previously (15) and as described in the legend to Fig. 8. Murine primary arterial smooth muscle cultures were prepared and documented to represent arterial smooth muscle cells as described previously (16).

Bottom Line: Adenoviral delivery of hepatocyte growth factor (HGF) to rodent islets improves islet graft survival and function, markedly reducing the number of islets required to achieve glucose control.Unilateral nephrectomy resulted in an immediate return of glucose to baseline diabetic levels.Interestingly, adenoviral DNA, as well as mouse HGF (mHGF) mRNA derived from the adenovirus, were present for 42 days posttransplantation.

View Article: PubMed Central - PubMed

Affiliation: Division of Endocrinology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA. taeschn@dom.pitt.edu

ABSTRACT

Objective: Adenoviral delivery of hepatocyte growth factor (HGF) to rodent islets improves islet graft survival and function, markedly reducing the number of islets required to achieve glucose control. Here, we asked whether these prior observations in rodent models extend to nonhuman primate (NHP) islets.

Research design and methods: NHP islets were transduced with murine (Ad.mHGF) or human (Ad.hHGF) adenoviral HGF (Ad.HGF) at low multiplicity of infection and studied in vitro. To study the function of Ad.HGF-transduced NHP islets in vivo, a renal subcapsular marginal mass islet transplant model was developed in streptozotocin-induced diabetic NOD-SCID mice.

Results: Baseline glucose values were 454.7 +/- 11.3 mg/dl (n = 7). Transplant of 500 NHP islet equivalents (IE) had only a marginal effect on blood glucose (369.1 +/- 9.7 mg/dl, n = 5). In striking contrast, 500 NHP IE transduced with Ad.mHGF promptly and continuously corrected blood glucose (142.0 +/- 6.2 mg/dl, n = 7) for the 6-week duration of the experiment. Unilateral nephrectomy resulted in an immediate return of glucose to baseline diabetic levels. Interestingly, adenoviral DNA, as well as mouse HGF (mHGF) mRNA derived from the adenovirus, were present for 42 days posttransplantation. Surprisingly, transplant of 500 IE with Ad.hHGF, as compared with Ad.mHGF, resulted in only marginal correction of blood glucose, suggesting that human HGF is less efficient than mHGF in this system.

Conclusions: These studies demonstrate that mHGF markedly improves islet transplant outcomes in the highest preclinical species examined to date. HGF has promise as an agent that can improve islet mass and function in transplant models and likely in other models of types 1 and 2 diabetes.

Show MeSH
Related in: MedlinePlus