INGAP protein involved in pancreatic islet neogenesis

a technology of pancreatic islet and ingap protein, which is applied in the direction of acute pancreatitis-associated proteins, antibody medical ingredients, peptide sources, etc., can solve the problems of limited regeneration capacity and limited regeneration capacity, and achieve the effect of enhancing the survival rate or survival time and enhancing the life span of pancreatic islet cells

Inactive Publication Date: 2006-09-19
EASTERN VIRGINIA MEDICAL SCHOOL +1
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  • Summary
  • Abstract
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Benefits of technology

[0057]According to another embodiment of the invention a method of enhancing the life span of pancreatic islet cells encapsulated in a polycarbon shell is provided. The method...

Problems solved by technology

However, they have a limited capacity for regeneration.
This limite...

Method used

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  • INGAP protein involved in pancreatic islet neogenesis
  • INGAP protein involved in pancreatic islet neogenesis
  • INGAP protein involved in pancreatic islet neogenesis

Examples

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example 1

[0119]This example describes the cloning and isolation of a cDNA encoding a novel, developmentally regulated, pancreatic protein.

[0120]We hypothesized that a unique locally produced factor(s) is responsible for islet cell regeneration. Using the recently developed mRNA differential display technique (5,6) to compare genes differentially expressed in cellophane wrapped (CW) versus control pancreata (CP) allowed us to identify a cDNA clone (RD19-2) which was uniquely expressed in cellophane wrapped pancreas.

[0121]A cDNA library was constructed from mRNA isolated from cellophane wrapped hamster pancreas using oligo d(T) primed synthesis, and ligation into pcDNA3 vector (Invitrogen). The number of primary recombinants in the library was 1.2×104 with an average size of 1.1 kb. The cDNA library was screened for clones of interest using high density colony plating techniques. Colonies were lifted onto nylon membranes (Schleicher & Schnell) and further digested with proteinase K (50(g / ml). ...

example 2

[0122]This example compares the sequence of INGAP to other proteins with which it shares homology.

[0123]The nucleotide sequence of the hamster INGAP clone with the longest cDNA insert was determined. As shown in FIGS. 1A and 1B the hamster cDNA comprises 747 nucleotides (nt), exclusive of the poly(A) tail and contains a major open reading frame encoding a 175 amino acid protein. The open reading frame is followed by a 3′-untranslated region of 206 nt. A typical polyadenylation signal is present 11 Nt upstream of the poly(A) tail. The predicted INGAP protein shows structural homology to both the PAP / HIP family of genes which is associated with pancreatitis or liver adenocarcinoma (7-11) and the Reg / PSP / lithostatine family of genes (13,15) which has been shown to stimulate pancreatic beta-cell growth (14) and might play a role in pancreatic islet regeneration. Comparison of the nucleotide sequence and their deduced amino acids between hamster INGAP and rat PAP-I shows a high degree of...

example 3

[0124]This example demonstrates the temporal expression pattern of INGAP upon cellophane-wrapping.

[0125]In order to determine the temporal expression of the INGAP gene, total RNA extracted from CP and CW pancreas was probed with the hamster INGAP cDNA clone in Northern blot analysis. A strong single transcript of 900 bp was detected (FIGS. 3A, 3B and 3C) 1 and 2 days after cellophane wrapping which disappeared by 6 through 42 days and was absent from CP. INGAP mRNA is associated with CW induced pancreatic islet neogenesis, since it is present only after CW. It is not likely that the increased expression of INGAP is associated with acute pancreatitis as is the case with the PAP family of genes. During the acute phase of pancreatitis the concentrations of most mRNAs encoding pancreatic enzymes including amylase are decreased significantly (16,18). In contrast, in the CW model of islet neogenesis in which high expression of INGAP has been detected, amylase gene expression was simultane...

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Abstract

Cellophane wrapping (CW) of hamster pancreas induces proliferation of duct epithelial cells followed by endocrine cell differentiation and islet neogenesis. Using the mRNA differential display technique a cDNA clone expressed in cellophane wrapped but not in control pancreata was identified. Using this cDNA as a probe, a cDNA library was screened and a gene not previously described was identified and named INGAP.

Description

[0001]This application is a reissue application of U.S. Pat. No. 5,834,590, filed as application Ser. No. 08 / 401,530 on Feb. 22, 1995.BACKGROUND OF THE INVENTION[0002]Pancreatic islets of Langerhans are the only organ of insulin production in the body. However, they have a limited capacity for regeneration. This limited regeneration capacity predisposes mammals to develop diabetes mellitus. Thus there is a need in the art of endocrinology for products which can stimulate the regeneration of islets of Langerhans to prevent or ameliorate the symptoms of diabetes mellitus.[0003]One model of pancreatic islet cell regeneration involves cellophane-wrapping of the pancreas in the Syrian golden hamster (1). Wrapping of the pancreas induces the formation of new endocrine cells which appear to arise from duct epithelium (2-4). There is a need in the art to identify and isolate the factor(s) which is responsible for islet cell regeneration.SUMMARY OF THE INVENTION[0004]It is an object of the i...

Claims

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Application Information

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IPC IPC(8): C07K14/00A61K38/00C07K14/47
CPCA01K2217/05A61K38/00C07K14/4733C07K14/474C07K2319/00
Inventor VINIK, AARON I.PITTENGER, GARY L.RAFAELOFF-PHAIL, RONITROSENBERG, LAWRENCE
Owner EASTERN VIRGINIA MEDICAL SCHOOL
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