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A therapeutic gene cocktail for heart regeneration

A heart regeneration, heart technology, applied in gene therapy, genetic engineering, medical preparations containing active ingredients, etc., can solve the problem of not understanding the detailed mechanism or function of the cell cycle

Active Publication Date: 2018-03-27
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Furthermore, the detailed mechanism or function of CM re-entry into the cell cycle at different time points is still not understood

Method used

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  • A therapeutic gene cocktail for heart regeneration
  • A therapeutic gene cocktail for heart regeneration
  • A therapeutic gene cocktail for heart regeneration

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0119] Example 1: Therapeutic Gene Mixtures for Cardiac Regeneration

[0120] Materials and methods

[0121] (i) Isolation of cardiomyocytes and non-cardiomyocytes from neonatal mouse hearts

[0122] Cardiomyocytes and non-cardiomyocytes were isolated from 2- or 3-day-old C57BL / 6 mice and cultured according to the following protocol: (Condorelli, Morisco et al. FASEB J. 16(13):1732-1737; 2002). Briefly, after removal of the atrium and aorta, heart tissue was dissected and dissociated with 1 mg / ml of trypsin (Trypsin, Sigma-Aldrich) at 4°C for 2 hours. Add 0.8 mg / ml of collagenase II (Collagenase II, Invitrogen), react at 37° C. for 15 minutes, filter with a 40-micron filter to remove cell debris, and collect the cells.

[0123] Stain with mitochondrial dye-tetramethylrhodamine methyl acetate (methyl ester perchlorate, TMRM, LifeTechnology), and use fluorescence activated cell separator FACS (Hattori, et al., Nat Methods 7(1):61- 66; 2010) analyzed and isolated the cell popu...

Embodiment 2

[0161] Example 2: Specific gene mixture preparation can significantly improve the proliferation of cardiomyocytes in adult rats

[0162] Materials and methods

[0163] (i) Cardiomyocytes isolated from adult mice

[0164] Ventricular CMs of adult mice were isolated from male mice on the Langendorff apparatus. After 10 minutes of heparinization, the heart was removed from the anesthetized mouse and cannulated using calcium-free (Ca2 + ) benchtop solution (Tyrodesolution, 120.4 mmol per liter of NaCl, 14.7 mmol per liter of KCl, 0.6 mmol per liter of KH 2 PO 4 , 0.6 mmol Na per liter 2 HPO 4 , 1.2 mmol of MgSO per liter 4 , 1.2 mmol per liter of HEPES, 4.6 mmol per liter of NaHCO 3 , 30 millimoles per liter of taurine (Taurine), per liter of 10 millimoles of BDM, per liter of 5.5 millimoles of glucose) for reverse perfusion. After perfusion for 3 minutes, mix with calcium ion-free benchtop solution (containing collagenase B (0.4 mg per gram of body weight, Roche), collage...

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Abstract

A combination of active agents selected from a FoxM1 enhancer, an Id1 enhancer, and a JNK3 inhibitor and the uses thereof in promoting cardiomyocyte proliferation and treating heart diseases in a subject in need of the treatment.

Description

[0001] This application claims priority under 35 U.S.C. §119(e) to U.S. Provisional Application No. 62 / 149,854, "A THERAPEUTICGENE COCKTAIL FOR HEART REGENERATION," filed April 20, 2015, which is incorporated by reference in its entirety . Background technique [0002] Cardiomyocyte (CM) proliferative ability of mature individuals in mammals is poor, so heart regeneration has been a long-standing challenge in this field. CM will undergo the last DNA synthesis and mitosis within one week of birth, and will no longer enter the cell cycle after becoming a binuclear cell. Although the remaining CM still has a certain degree of proliferative ability after the heart is damaged, its proliferative efficiency is extremely low (Naqvi et al., Cell 157(4):795-807; 2014). [0003] It is known that somatic cells can be reprogrammed to return to a state of pluripotent cells similar to embryonic stem cells (ESCs) with high proliferative capacity (Takahashi et al., Cell126(4):663 -676; 2006)...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/17A61P9/00A61K48/00A61K31/713
CPCA61K31/713A61K45/06A61K38/1709C12N2710/10343A61K48/005A61P9/10A61K2300/00C12N2310/531
Inventor 谢清河郑媛元
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