Engineered M13 Peptide Carrier Promotes Angiogenic Potential of Patient-Derived Human Cardiac Progenitor Cells and In Vivo Engraftment

장웅비,지승택,박지혜,Kim Yeon-Ju,Kang Songhwa,김다연,이나경,김진수,Lim Hye Ji,최재우,LE THI HONG VAN,LY THANH TRUONG GIANG,비누스,김동환,하종성,윤지수,Baek Sang Hong,권상모 한국조직공학과 재생의학회 2020 조직공학과 재생의학 Vol.17 No.3

BACKGROUND: Despite promising advances in stem cell-based therapy, the treatment of ischemic cardiovascular diseases remains a big challenge due to both the insufficient in vivo viability of transplanted cells and poor angiogenic potential of stem cells. The goal of this study was to develop therapeutic human cardiac progenitor cells (hCPCs) for ischemic cardiovascular diseases with a novel M13 peptide carrier. METHOD: In this study, an engineered M13 peptide carrier was successfully generated using a QuikChange Kit. The cellular function of M13 peptide carrier-treated hCPCs was assessed using a tube formation assay and scratch wound healing assay. The in vivo engraftment and cell survival bioactivities of transplanted cells were demonstrated by immunohistochemistry after hCPC transplantation into a myocardial infarction animal model. RESULTS: The engineered M13RGD?SDKP peptide carrier, which expressed RGD peptide on PIII site and SDKP peptide on PVIII site, did not affect morphologic change and proliferation ability in hCPCs. In contrast, hCPCs treated with M13RGD?SDKP showed enhanced angiogenic capacity, including tube formation and migration capacity. Moreover, transplanted hCPCs with M13RGD?SDKP were engrafted into the ischemic region and promoted in vivo cell survival. CONCLUSION: Our present data provides a promising protocol for CPC-based cell therapy via short-term cell priming of hCPCs with engineered M13RGD?SDKP before cell transplantation for treatment of cardiovascular disease.

Engineered M13 Nanofiber Accelerates Ischemic Neovascularization by Enhancing Endothelial Progenitor Cells

이준희,김성욱,지승택,김연주,장웅비,오진우,김재호,유소영,백상홍,권상모 한국조직공학과 재생의학회 2017 조직공학과 재생의학 Vol.14 No.6

Dysfunction or loss of blood vessel causes several ischemic diseases.Although endothelial progenitor cells (EPCs) are a promising source for cell-based therapy, ischemia-induced pathophysiological condition limits the recovery rate by causing drastic cell death. To overcome this issue, we attempted to develop a cell-targeted peptide delivery and priming system to enhance EPCbased neovascularization using an engineered M13 bacteriophage harboring nanofibrous tubes displaying *2700 multiple functional motifs. The M13 nanofiber was modified by displaying RGD, which is an integrin-docking peptide, on the minor coat protein, and bymutilayering SDKPmotifs,which are the key active sites for thymosin b4, on themajor coat protein. The engineered M13 nanofiber dramatically enhanced ischemic neovascularization by activating intracellular and extracellular processes such as proliferation, migration, and tube formation in the EPCs. Furthermore, transplantation of the primed EPCs with the M13 nanofiber harboring RGD and SDKP facilitated functional recovery and neovascularization in a murine hindlimb ischemia model. Overall, this study demonstrates the effectiveness of theM13 nanofiber-based novel peptide deliveryandprimingstrategy inpromotingEPC bioactivity and neovessel regeneration. To our knowledge, this is first report onM13 nanofibers harboring dual functional motifs, the use of which might be a novel strategy for stem and progenitor cell therapy against cardiovascular ischemic diseases.

근해 대형선망어업의 온실가스 배출량 정량적 분석

신동원(Dongwon SHIN),장웅비(Wungbi JANG),이지훈(Jihoon LEE) 전남대학교 어업기술연구소 2016 어업기술연구소보고지 Vol.9 No.1

Ozone layer depletion and global warming related to GHG (greenhouse gases) emissions from industries are a major issue globally. As these efforts, The parties of the Kyoto protocol adopted in the 3th UNFCCC’s conference set targets for average 5.2 percent reduction of GHG emissions from 1990 until 2012, should apply greenhouse gas emissions trading. The 18th UNFCCC’s conference of the parties to be held in Doha, Qatar agreed the Doha amendment to extend the Kyoto protocol that expires in 2012 until 2020. Furthermore, GHG emissions from the fishery industries also represent an important issue, as indicated by Responsible Fisheries at Cancun, Mexico, in The 16th UNFCCC’s conference of the parties, United nations conference on environment & development accepted Responsible Fisheries as important concern area. However, few research on the GHG emissions from Korean fisheries have been performed. Therefore, a quantitative analysis of GHG emissions from the major Korean fisheries in needed before guidelines for reducing GHG emissions from the fishing industry can be established. The aim of this study was to assess the present GHG emissions from the Korean offshore large purse seine fishery using the Life Cycle Assessment (LCA) method quantitatively. The result of this study will be helpful to establish a reducing method of GHG emissions.

Long-Term Priming by Three Small Molecules Is a Promising Strategy for Enhancing Late Endothelial Progenitor Cell Bioactivities

김연주,지승택,김다연,정석윤,강송화,박지혜,장웅비,윤지수,하종성,이동형,권상모 한국분자세포생물학회 2018 Molecules and cells Vol.41 No.6

Endothelial progenitor cells (EPCs) and outgrowth endothelial cells (OECs) play a pivotal role in vascular regeneration in ischemic tissues; however, their therapeutic application in clinical settings is limited due to the low quality and quantity of patient-derived circulating EPCs. To solve this problem, we evaluated whether three priming small molecules (tauroursodeoxycholic acid, fucoidan, and oleuropein) could enhance the angiogenic potential of EPCs. Such enhancement would promote the cellular bioactivities and help to develop functionally improved EPC therapeutics for ischemic diseases by accelerating the priming effect of the defined physiologi-cal molecules. We found that preconditioning of each of the three small molecules significantly induced the differentiation potential of CD34+ stem cells into EPC lineage cells. Notably, long-term priming of OECs with the three chemical cocktail (OEC-3C) increased the pro-liferation potential of EPCs via ERK activation. The migration, invasion, and tube-forming capacities were also significantly enhanced in OEC-3Cs compared with unprimed OECs. Further, the cell survival ratio was dra-matically increased in OEC-3Cs against H2O2-induced oxidative stress via the augmented expression of Bcl-2, a pro-survival protein. In conclusion, we identified three small molecules for enhancing the bioactivities of ex vivo-expanded OECs for vascular repair. Long-term 3C priming might be a promising methodology for EPC-based therapy against ischemic diseases.

Human cardiac stem cells rejuvenated by modulating autophagy with MHY-1685 enhance the therapeutic potential for cardiac repair

Park Ji Hye,Kim Hyeok,Moon Hyung Ryong,박봉우,Park Jae-Hyun,Sim Woo-Sup,Kim Jin-Ju,Lim Hye Ji,Kim Yeon-Ju,지승택,장웅비,Rethineswaran Vinoth Kumar,Van Le Thi Hong,Giang Ly Thanh Truong,Yun Jisoo,Ha Jong Seong 생화학분자생물학회 2021 Experimental and molecular medicine Vol.53 No.-

Stem cell-based therapies with clinical applications require millions of cells. Therefore, repeated subculture is essential for cellular expansion, which is often complicated by replicative senescence. Cellular senescence contributes to reduced stem cell regenerative potential as it inhibits stem cell proliferation and differentiation as well as the activation of the senescence-associated secretory phenotype (SASP). In this study, we employed MHY-1685, a novel mammalian target of rapamycin (mTOR) inhibitor, and examined its long-term priming effect on the activities of senile human cardiac stem cells (hCSCs) and the functional benefits of primed hCSCs after transplantation. In vitro experiments showed that the MHY-1685‒primed hCSCs exhibited higher viability in response to oxidative stress and an enhanced proliferation potential compared to that of the unprimed senile hCSCs. Interestingly, priming MHY-1685 enhanced the expression of stemness-related markers in senile hCSCs and provided the differentiation potential of hCSCs into vascular lineages. In vivo experiment with echocardiography showed that transplantation of MHY-1685‒primed hCSCs improved cardiac function than that of the unprimed senile hCSCs at 4 weeks post-MI. In addition, hearts transplanted with MHY-1685-primed hCSCs exhibited significantly lower cardiac fibrosis and higher capillary density than that of the unprimed senile hCSCs. In confocal fluorescence imaging, MHY-1685‒primed hCSCs survived for longer durations than that of the unprimed senile hCSCs and had a higher potential to differentiate into endothelial cells (ECs) within the infarcted hearts. These findings suggest that MHY-1685 can rejuvenate senile hCSCs by modulating autophagy and that as a senescence inhibitor, MHY-1685 can provide opportunities to improve hCSC-based myocardial regeneration.