Direct reprogramming induces vascular regeneration post muscle ischemic injury

Keerat Kaur; Yoav Hadas; Ann Anu Kurian; Magdalena M. Żak; Jimeen Yoo; Asharee Mahmood; Hanna Girard; Rinat Komargodski; Toshiro Io; Maria Paola Santini; Nishat Sultana; Mohammad Tofael Kabir Sharkar; Ajit Magadum; Anthony Fargnoli; Seonghun Yoon; Elena Chepurko; Vadim Chepurko; Efrat Eliyahu; Dalila Pinto; Djamel Lebeche; Jason C. Kovacic; Roger J. Hajjar; Shahin Rafii; Lior Zangi

doi:10.1016/j.ymthe.2021.07.014

Direct reprogramming induces vascular regeneration post muscle ischemic injury

Keerat Kaur, Yoav Hadas, Ann Anu Kurian, Magdalena M. Żak, Jimeen Yoo, Asharee Mahmood, Hanna Girard, Rinat Komargodski, Toshiro Io, Maria Paola Santini, Nishat Sultana, Mohammad Tofael Kabir Sharkar, Ajit Magadum, Anthony Fargnoli, Seonghun Yoon, Elena Chepurko, Vadim Chepurko, Efrat Eliyahu, Dalila Pinto, Djamel LebecheJason C. Kovacic, Roger J. Hajjar, Shahin Rafii, Lior Zangi^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

13 Scopus citations

Abstract

Reprogramming non-cardiomyocytes (non-CMs) into cardiomyocyte (CM)-like cells is a promising strategy for cardiac regeneration in conditions such as ischemic heart disease. Here, we used a modified mRNA (modRNA) gene delivery platform to deliver a cocktail, termed 7G-modRNA, of four cardiac-reprogramming genes—Gata4 (G), Mef2c (M), Tbx5 (T), and Hand2 (H)—together with three reprogramming-helper genes—dominant-negative (DN)-TGFβ, DN-Wnt8a, and acid ceramidase (AC)—to induce CM-like cells. We showed that 7G-modRNA reprogrammed 57% of CM-like cells in vitro. Through a lineage-tracing model, we determined that delivering the 7G-modRNA cocktail at the time of myocardial infarction reprogrammed ∼25% of CM-like cells in the scar area and significantly improved cardiac function, scar size, long-term survival, and capillary density. Mechanistically, we determined that while 7G-modRNA cannot create de novo beating CMs in vitro or in vivo, it can significantly upregulate pro-angiogenic mesenchymal stromal cells markers and transcription factors. We also demonstrated that our 7G-modRNA cocktail leads to neovascularization in ischemic-limb injury, indicating CM-like cells importance in other organs besides the heart. modRNA is currently being used around the globe for vaccination against COVID-19, and this study proves this is a safe, highly efficient gene delivery approach with therapeutic potential to treat ischemic diseases.

Original language	American English
Pages (from-to)	3042-3058
Number of pages	17
Journal	Molecular Therapy
Volume	29
Issue number	10
DOIs	https://doi.org/10.1016/j.ymthe.2021.07.014
State	Published - 6 Oct 2021
Externally published	Yes

Bibliographical note

Funding Information:
This work was mostly funded by a sponsor agreement given by Ono Pharmaceutical Co., Ltd. (Osaka, Japan) to the Zangi lab. It was also partially funded by a cardiology start-up grant awarded to the Zangi laboratory and by NIH grants R01 HL142768-01 and R01 HL149137-01 . Y.H. and D.P. were supported by grants of the National Institute of Mental Health (NIMH) ( R01-MH109715 , D.P. and R21-MH105881 , D.P).

Funding Information:
The authors acknowledge Okino Tomotaka, Koji Shinozaki, Tomoyuki Bando, Tetsuya Sugiyama, and Matsushita Yuichiro for their help with this manuscript. This work was mostly funded by a sponsor agreement given by Ono Pharmaceutical Co. Ltd. (Osaka, Japan) to the Zangi lab. It was also partially funded by a cardiology start-up grant awarded to the Zangi laboratory and by NIH grants R01 HL142768-01 and R01 HL149137-01. Y.H. and D.P. were supported by grants of the National Institute of Mental Health (NIMH) (R01-MH109715, D.P. and R21-MH105881, D.P). L.Z. and K.K. are inventors of a Patent Cooperation Treaty application WO2021050877A1 (Compositions including molecules of modified mRNA and methods of using the same), which covers the results in this manuscript.

Publisher Copyright:
© 2021 The Author(s)

Keywords

cardiac repair
cardiac reprogramming
cardiovascular reprogramming
gene therapy
hindlimb ischemia
modified mRNA

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1016/j.ymthe.2021.07.014

Cite this

Kaur, K., Hadas, Y., Kurian, A. A., Żak, M. M., Yoo, J., Mahmood, A., Girard, H., Komargodski, R., Io, T., Santini, M. P., Sultana, N., Sharkar, M. T. K., Magadum, A., Fargnoli, A., Yoon, S., Chepurko, E., Chepurko, V., Eliyahu, E., Pinto, D., ... Zangi, L. (2021). Direct reprogramming induces vascular regeneration post muscle ischemic injury. Molecular Therapy, 29(10), 3042-3058. https://doi.org/10.1016/j.ymthe.2021.07.014

@article{3e6c79a04bee4ee8920d421df1c9d4c4,

title = "Direct reprogramming induces vascular regeneration post muscle ischemic injury",

abstract = "Reprogramming non-cardiomyocytes (non-CMs) into cardiomyocyte (CM)-like cells is a promising strategy for cardiac regeneration in conditions such as ischemic heart disease. Here, we used a modified mRNA (modRNA) gene delivery platform to deliver a cocktail, termed 7G-modRNA, of four cardiac-reprogramming genes—Gata4 (G), Mef2c (M), Tbx5 (T), and Hand2 (H)—together with three reprogramming-helper genes—dominant-negative (DN)-TGFβ, DN-Wnt8a, and acid ceramidase (AC)—to induce CM-like cells. We showed that 7G-modRNA reprogrammed 57% of CM-like cells in vitro. Through a lineage-tracing model, we determined that delivering the 7G-modRNA cocktail at the time of myocardial infarction reprogrammed ∼25% of CM-like cells in the scar area and significantly improved cardiac function, scar size, long-term survival, and capillary density. Mechanistically, we determined that while 7G-modRNA cannot create de novo beating CMs in vitro or in vivo, it can significantly upregulate pro-angiogenic mesenchymal stromal cells markers and transcription factors. We also demonstrated that our 7G-modRNA cocktail leads to neovascularization in ischemic-limb injury, indicating CM-like cells importance in other organs besides the heart. modRNA is currently being used around the globe for vaccination against COVID-19, and this study proves this is a safe, highly efficient gene delivery approach with therapeutic potential to treat ischemic diseases.",

keywords = "cardiac repair, cardiac reprogramming, cardiovascular reprogramming, gene therapy, hindlimb ischemia, modified mRNA",

author = "Keerat Kaur and Yoav Hadas and Kurian, {Ann Anu} and {\.Z}ak, {Magdalena M.} and Jimeen Yoo and Asharee Mahmood and Hanna Girard and Rinat Komargodski and Toshiro Io and Santini, {Maria Paola} and Nishat Sultana and Sharkar, {Mohammad Tofael Kabir} and Ajit Magadum and Anthony Fargnoli and Seonghun Yoon and Elena Chepurko and Vadim Chepurko and Efrat Eliyahu and Dalila Pinto and Djamel Lebeche and Kovacic, {Jason C.} and Hajjar, {Roger J.} and Shahin Rafii and Lior Zangi",

note = "Funding Information: This work was mostly funded by a sponsor agreement given by Ono Pharmaceutical Co., Ltd. (Osaka, Japan) to the Zangi lab. It was also partially funded by a cardiology start-up grant awarded to the Zangi laboratory and by NIH grants R01 HL142768-01 and R01 HL149137-01 . Y.H. and D.P. were supported by grants of the National Institute of Mental Health (NIMH) ( R01-MH109715 , D.P. and R21-MH105881 , D.P). Funding Information: The authors acknowledge Okino Tomotaka, Koji Shinozaki, Tomoyuki Bando, Tetsuya Sugiyama, and Matsushita Yuichiro for their help with this manuscript. This work was mostly funded by a sponsor agreement given by Ono Pharmaceutical Co. Ltd. (Osaka, Japan) to the Zangi lab. It was also partially funded by a cardiology start-up grant awarded to the Zangi laboratory and by NIH grants R01 HL142768-01 and R01 HL149137-01. Y.H. and D.P. were supported by grants of the National Institute of Mental Health (NIMH) (R01-MH109715, D.P. and R21-MH105881, D.P). L.Z. and K.K. are inventors of a Patent Cooperation Treaty application WO2021050877A1 (Compositions including molecules of modified mRNA and methods of using the same), which covers the results in this manuscript. Publisher Copyright: {\textcopyright} 2021 The Author(s)",

year = "2021",

month = oct,

day = "6",

doi = "10.1016/j.ymthe.2021.07.014",

language = "American English",

volume = "29",

pages = "3042--3058",

journal = "Molecular Therapy",

issn = "1525-0016",

publisher = "Cell Press",

number = "10",

}

Kaur, K, Hadas, Y, Kurian, AA, Żak, MM, Yoo, J, Mahmood, A, Girard, H, Komargodski, R, Io, T, Santini, MP, Sultana, N, Sharkar, MTK, Magadum, A, Fargnoli, A, Yoon, S, Chepurko, E, Chepurko, V, Eliyahu, E, Pinto, D, Lebeche, D, Kovacic, JC, Hajjar, RJ, Rafii, S & Zangi, L 2021, 'Direct reprogramming induces vascular regeneration post muscle ischemic injury', Molecular Therapy, vol. 29, no. 10, pp. 3042-3058. https://doi.org/10.1016/j.ymthe.2021.07.014

TY - JOUR

T1 - Direct reprogramming induces vascular regeneration post muscle ischemic injury

AU - Kaur, Keerat

AU - Hadas, Yoav

AU - Kurian, Ann Anu

AU - Żak, Magdalena M.

AU - Yoo, Jimeen

AU - Mahmood, Asharee

AU - Girard, Hanna

AU - Komargodski, Rinat

AU - Io, Toshiro

AU - Santini, Maria Paola

AU - Sultana, Nishat

AU - Sharkar, Mohammad Tofael Kabir

AU - Magadum, Ajit

AU - Fargnoli, Anthony

AU - Yoon, Seonghun

AU - Chepurko, Elena

AU - Chepurko, Vadim

AU - Eliyahu, Efrat

AU - Pinto, Dalila

AU - Lebeche, Djamel

AU - Kovacic, Jason C.

AU - Hajjar, Roger J.

AU - Rafii, Shahin

AU - Zangi, Lior

N1 - Funding Information: This work was mostly funded by a sponsor agreement given by Ono Pharmaceutical Co., Ltd. (Osaka, Japan) to the Zangi lab. It was also partially funded by a cardiology start-up grant awarded to the Zangi laboratory and by NIH grants R01 HL142768-01 and R01 HL149137-01 . Y.H. and D.P. were supported by grants of the National Institute of Mental Health (NIMH) ( R01-MH109715 , D.P. and R21-MH105881 , D.P). Funding Information: The authors acknowledge Okino Tomotaka, Koji Shinozaki, Tomoyuki Bando, Tetsuya Sugiyama, and Matsushita Yuichiro for their help with this manuscript. This work was mostly funded by a sponsor agreement given by Ono Pharmaceutical Co. Ltd. (Osaka, Japan) to the Zangi lab. It was also partially funded by a cardiology start-up grant awarded to the Zangi laboratory and by NIH grants R01 HL142768-01 and R01 HL149137-01. Y.H. and D.P. were supported by grants of the National Institute of Mental Health (NIMH) (R01-MH109715, D.P. and R21-MH105881, D.P). L.Z. and K.K. are inventors of a Patent Cooperation Treaty application WO2021050877A1 (Compositions including molecules of modified mRNA and methods of using the same), which covers the results in this manuscript. Publisher Copyright: © 2021 The Author(s)

PY - 2021/10/6

Y1 - 2021/10/6

N2 - Reprogramming non-cardiomyocytes (non-CMs) into cardiomyocyte (CM)-like cells is a promising strategy for cardiac regeneration in conditions such as ischemic heart disease. Here, we used a modified mRNA (modRNA) gene delivery platform to deliver a cocktail, termed 7G-modRNA, of four cardiac-reprogramming genes—Gata4 (G), Mef2c (M), Tbx5 (T), and Hand2 (H)—together with three reprogramming-helper genes—dominant-negative (DN)-TGFβ, DN-Wnt8a, and acid ceramidase (AC)—to induce CM-like cells. We showed that 7G-modRNA reprogrammed 57% of CM-like cells in vitro. Through a lineage-tracing model, we determined that delivering the 7G-modRNA cocktail at the time of myocardial infarction reprogrammed ∼25% of CM-like cells in the scar area and significantly improved cardiac function, scar size, long-term survival, and capillary density. Mechanistically, we determined that while 7G-modRNA cannot create de novo beating CMs in vitro or in vivo, it can significantly upregulate pro-angiogenic mesenchymal stromal cells markers and transcription factors. We also demonstrated that our 7G-modRNA cocktail leads to neovascularization in ischemic-limb injury, indicating CM-like cells importance in other organs besides the heart. modRNA is currently being used around the globe for vaccination against COVID-19, and this study proves this is a safe, highly efficient gene delivery approach with therapeutic potential to treat ischemic diseases.

AB - Reprogramming non-cardiomyocytes (non-CMs) into cardiomyocyte (CM)-like cells is a promising strategy for cardiac regeneration in conditions such as ischemic heart disease. Here, we used a modified mRNA (modRNA) gene delivery platform to deliver a cocktail, termed 7G-modRNA, of four cardiac-reprogramming genes—Gata4 (G), Mef2c (M), Tbx5 (T), and Hand2 (H)—together with three reprogramming-helper genes—dominant-negative (DN)-TGFβ, DN-Wnt8a, and acid ceramidase (AC)—to induce CM-like cells. We showed that 7G-modRNA reprogrammed 57% of CM-like cells in vitro. Through a lineage-tracing model, we determined that delivering the 7G-modRNA cocktail at the time of myocardial infarction reprogrammed ∼25% of CM-like cells in the scar area and significantly improved cardiac function, scar size, long-term survival, and capillary density. Mechanistically, we determined that while 7G-modRNA cannot create de novo beating CMs in vitro or in vivo, it can significantly upregulate pro-angiogenic mesenchymal stromal cells markers and transcription factors. We also demonstrated that our 7G-modRNA cocktail leads to neovascularization in ischemic-limb injury, indicating CM-like cells importance in other organs besides the heart. modRNA is currently being used around the globe for vaccination against COVID-19, and this study proves this is a safe, highly efficient gene delivery approach with therapeutic potential to treat ischemic diseases.

KW - cardiac repair

KW - cardiac reprogramming

KW - cardiovascular reprogramming

KW - gene therapy

KW - hindlimb ischemia

KW - modified mRNA

UR - http://www.scopus.com/inward/record.url?scp=85117120150&partnerID=8YFLogxK

U2 - 10.1016/j.ymthe.2021.07.014

DO - 10.1016/j.ymthe.2021.07.014

M3 - Article

C2 - 34332145

AN - SCOPUS:85117120150

SN - 1525-0016

VL - 29

SP - 3042

EP - 3058

JO - Molecular Therapy

JF - Molecular Therapy

IS - 10

ER -

Direct reprogramming induces vascular regeneration post muscle ischemic injury

Abstract

Bibliographical note

Keywords

UN SDGs

Access to Document

Other files and links

Fingerprint

Cite this