Patent NZ-592821-A (NZ)
Title: Regeneration of pancreatic islets and reversal of diabetes by islet transcription factor genes delivered in vivo
Inventor: GRAYBURN PAUL A
IPC: C12N5/071
Filed: 2009-11-13
Abstract: Disclosed is a composition for ultrasound-targeted microbubble destruction in the pancreas comprising a pre-assembled liposome-nucleic acid microbubble complex, wherein the pre-assembled liposome-nucleic acid complex comprises a nucleic acid comprising a Neuro D gene under the control of an insulin promoter, wherein disruption of the microbubble in the pancreas at a target site delivers the nucleic acid into pancreas cells at the location of the ultrasound disruption, wherein cells that incorporate the nucleic acid express insulin in response to high blood glucose levels. Further disclosed is a method of making an isolated target cell insulin responsive comprising providing a nucleic acid segment comprising a Neuro D gene under the control of an insulin responsive promoter selected form RIP4.1, CUB1, RIP2.1, RIP3.1 or HIP3.1 promoter, delivering the nucleic acid segment into the isolated target cell as a liposome-nucleic acid microbubble complex, and maintaining the target cell under conditions effective to express a insulin responsive regulatory gene; wherein expression of the Neuro D in the target cell causes the cell to respond to high blood glucose. Use of one or more insulin responsive regulatory genes operatively linked to an insulin promoter region in the manufacture of a medicament for the treatment of a diabetic patient by restoring insulin responsiveness wherein treatment comprises the steps of: obtaining an isolated nucleic acid segment comprising the one or more insulinresponsive regulatory genes operatively linked to a insulin promoter region comprising a genomic fragment of the insulin promoter comprising a 5' untranslated region, of exon1, intron1 and exon2 of the insulin gene; transferring the nucleic acid segment into a target cell as a liposome-nucleic acid microbubble complex; and maintaining the target cell under conditions effective to express the insulin responsive regulatory gene; wherein expression of the insulin responsive regulatory gene in the target cell causes the cell to respond to high blood glucose, wherein the one or more insulin responsive regulatory genes operatively linked to an insulin promoter region comprise Neuro D. Also disclosed is An isolated nucleic acid comprising an insulin promoter region comprising a genomic fragment of the insulin promoter comprising a 5' untranslated region, exon1, intron1 and exon2 of the insulin gene upstream from one or more insulin responsive genes, wherein the isolated nucleic acid is in the form of a liposome-nucleic acid microbubble complex. Also disclosed is An ex vivo cell made insulin responsive by a method comprising contacting the ex vivo cell with a pre-assembled liposome-nucleic acid microbubble complex, wherein the pre-assembled liposome-nucleic acid complex comprises a Neuro D gene under the control of an insulin promoter comprising a 5' untranslated region of exon1, intron1 and exon2 of the insulin gene, wherein disruption of the microbubble delivers the nucleic acid into the cell, wherein the cell expresses insulin in response to high blood glucose levels.