PIM1-minicircle as a therapeutic treatment for myocardial infarction.
PIM1, a pro-survival gene encoding a serine/ threonine kinase, influences cell proliferation and survival. Modification of cardiac progenitor cells (CPCs) or cardiomyocytes with PIM1 using a lentivirus-based delivery method showed long-term improved cardiac function after myocardial infarction (MI). However, lentivirus based delivery methods have stringent FDA regulation with respect to clinical trials. To provide an alternative and low risk PIM1 delivery method, this study examined the use of a non-viral modified plasmid-minicircle (MC) as a vehicle to deliver PIM1 into mouse CPCs (mCPCs) in vitro and the myocardium in vivo. MC containing a turbo gfp reporter gene (gfp-MC) was used as a transfection and injection control. PIM1 was subcloned into gfp-MC (PIM1-MC) and then transfected into mCPCs at an efficiency of 29.4±3.7%. PIM1-MC engineered mCPCs (PIM1-mCPCs) exhibit significantly (P<0.05) better survival rate under oxidative treatment. PIM1-mCPCs also exhibit 1.9±0.1 and 2.2±0.2 fold higher cell proliferation at 3 and 5 days post plating, respectively, as compared to gfp-MC transfected mCPCs control. PIM1-MC was injected directly into ten-week old adult FVB female mice hearts in the border zone immediately after MI. Delivery of PIM1 into myocardium was confirmed by GFP+ cardiomyocytes. Mice with PIM1-MC injection showed increased protection compared to gfp-MC injection groups measured by ejection fraction at 3 and 7 days post injury (P = 0.0379 and P = 0.0262 by t-test, respectively). Success of PIM1 delivery and integration into mCPCs in vitro and cardiomyocytes in vivo by MC highlights the possibility of a non-cell based therapeutic approach for treatment of ischemic heart disease and MI.
Biology Department, San Diego State University, San Diego, California, United States of America.
Nan
http://orcid.org/0000-0001-5194-2434
N
Liu
Biology Department, San Diego State University, San Diego, California, United States of America.
Bingyan J
BJ
Wang
Biology Department, San Diego State University, San Diego, California, United States of America.
Kathleen M
KM
Broughton
Biology Department, San Diego State University, San Diego, California, United States of America.
Roberto
R
Alvarez
Biology Department, San Diego State University, San Diego, California, United States of America.
Sailay
S
Siddiqi
Biology Department, San Diego State University, San Diego, California, United States of America.
Rebeca
R
Loaiza
Biology Department, San Diego State University, San Diego, California, United States of America.
Nicky
N
Nguyen
Biology Department, San Diego State University, San Diego, California, United States of America.
Pearl
P
Quijada
Biology Department, San Diego State University, San Diego, California, United States of America.
Natalie
N
Gude
Biology Department, San Diego State University, San Diego, California, United States of America.
Mark A
MA
Sussman
D000818:Animals; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D004195:Disease Models, Animal; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D051379:Mice; D032386:Myoblasts, Cardiac / Q000378:metabolism / Q000473:pathology; D009203:Myocardial Infarction / Q000235:genetics* / Q000503:physiopathology / Q000628:therapy*; D032383:Myocytes, Cardiac / Q000378:metabolism / Q000473:pathology; D010957:Plasmids / Q000235:genetics; D051573:Proto-Oncogene Proteins c-pim-1 / Q000235:genetics*; D014162:Transfection
C496636:Pim1 protein, mouse; D051573:Proto-Oncogene Proteins c-pim-1