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Article ; Online: Optimization of Synthesis of Modified mRNA.

Yoo, Jimeen / Zangi, Lior

Methods in molecular biology (Clifton, N.J.)

2022 Volume 2573, Page(s) 77–87

Abstract: Modified mRNA (modRNA) is a safe and effective vector for gene-based therapies. Notably, the safety of modRNA has been validated through COVID-19 vaccines which incorporate modRNA technology to translate spike proteins. Alternative gene delivery methods ... ...

Abstract	Modified mRNA (modRNA) is a safe and effective vector for gene-based therapies. Notably, the safety of modRNA has been validated through COVID-19 vaccines which incorporate modRNA technology to translate spike proteins. Alternative gene delivery methods using plasmids, lentiviruses, adenoviruses, and adeno-associated viruses have suffered from key challenges such as genome integration, delayed and uncontrolled expression, and immunogenic responses. However, modRNA poses no risk of genome integration, has transient and rapid expression, and lacks an immunogenic response. Our lab utilizes modRNA-based therapies to promote cardiac regeneration following myocardial infarction and heart failure. We have also developed and refined an optimized and economical method for synthesis of modRNA. Here, we provide an updated methodology with improved translational efficiency for in vitro and in vivo application.
MeSH term(s)	COVID-19/therapy ; COVID-19 Vaccines ; Gene Transfer Techniques ; Genetic Therapy/methods ; Humans ; RNA, Messenger/genetics ; RNA, Messenger/metabolism
Chemical Substances	COVID-19 Vaccines ; RNA, Messenger
Language	English
Publishing date	2022-08-30
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-2707-5_7
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Article: Lipid Nanoparticles for Organ-Specific mRNA Therapeutic Delivery.

Żak, Magdalena M / Zangi, Lior

Pharmaceutics

2021 Volume 13, Issue 10

Abstract: Advances in the using in vitro transcribed (IVT) modRNA in the past two decades, especially the tremendous recent success of mRNA vaccines against SARS-CoV-2, have brought increased attention to IVT mRNA technology. Despite its well-known use in ... ...

Abstract	Advances in the using in vitro transcribed (IVT) modRNA in the past two decades, especially the tremendous recent success of mRNA vaccines against SARS-CoV-2, have brought increased attention to IVT mRNA technology. Despite its well-known use in infectious disease vaccines, IVT modRNA technology is being investigated mainly in cancer immunotherapy and protein replacement therapy, with ongoing clinical trials in both areas. One of the main barriers to progressing mRNA therapeutics to the clinic is determining how to deliver mRNA to target cells and protect it from degradation. Over the years, many different vehicles have been developed to tackle this issue. Desirable vehicles must be safe, stable and preferably organ specific for successful mRNA delivery to clinically relevant cells and tissues. In this review we discuss various mRNA delivery platforms, with particular focus on attempts to create organ-specific vehicles for therapeutic mRNA delivery.
Language	English
Publishing date	2021-10-13
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2527217-2
ISSN	1999-4923
ISSN	1999-4923
DOI	10.3390/pharmaceutics13101675
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Article ; Online: Modified mRNA as a Therapeutic Tool for the Heart.

Kaur, Keerat / Zangi, Lior

Cardiovascular drugs and therapy

2020 Volume 34, Issue 6, Page(s) 871–880

Abstract: Despite various clinical modalities available for patients, heart disease remains among the leading causes of mortality and morbidity worldwide. Genetic medicine, particularly mRNA, has broad potential as a therapeutic. More specifically, mRNA-based ... ...

Abstract	Despite various clinical modalities available for patients, heart disease remains among the leading causes of mortality and morbidity worldwide. Genetic medicine, particularly mRNA, has broad potential as a therapeutic. More specifically, mRNA-based protein delivery has been used in the fields of cancer and vaccination, but recent changes to the structural composition of mRNA have led the scientific community to swiftly embrace it as a new drug to deliver missing genes to injured myocardium and many other organs. Modified mRNA (modRNA)-based gene delivery features transient but potent protein translation and low immunogenicity, with minimal risk of insertional mutagenesis. In this review, we compared and listed the advantages of modRNA over traditional vectors for cardiac therapy, with particular focus on using modRNA therapy in cardiac repair. We present a comprehensive overview of modRNA's role in cardiomyocyte (CM) proliferation, cardiac vascularization, and prevention of cardiac apoptosis. We also emphasize recent advances in modRNA delivery strategies and discuss the challenges for its clinical translation.
MeSH term(s)	Animals ; Cell Proliferation ; Gene Transfer Techniques ; Genetic Therapy/adverse effects ; Heart Diseases/genetics ; Heart Diseases/metabolism ; Heart Diseases/physiopathology ; Heart Diseases/therapy ; Humans ; Myocytes, Cardiac/metabolism ; Myocytes, Cardiac/pathology ; RNA, Messenger/adverse effects ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; RNA, Messenger/therapeutic use ; Recovery of Function ; Regeneration ; Risk Factors ; Treatment Outcome
Chemical Substances	RNA, Messenger
Keywords	covid19
Language	English
Publishing date	2020-08-21
Publishing country	United States
Document type	Journal Article ; Review
ZDB-ID	639068-7
ISSN	1573-7241 ; 0920-3206
ISSN (online)	1573-7241
ISSN	0920-3206
DOI	10.1007/s10557-020-07051-4
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Article ; Online: Lin28a cardiomyocyte-specific modified mRNA translation system induces cardiomyocyte cell division and cardiac repair.

Magadum, Ajit / Sun, Jiacheng / Singh, Neha / Kurian, Ann Anu / Chepurko, Elena / Fargnoli, Anthony / Hajjar, Roger / Zhang, Jianyi / Zangi, Lior

Journal of molecular and cellular cardiology

2024 Volume 188, Page(s) 61–64

Abstract: The mammalian heart has a limited regenerative capacity. Previous work suggested the heart can regenerate during development and immediately after birth by inducing cardiomyocyte (CM) proliferation; however, this capacity is lost seven days after birth. ... ...

Abstract	The mammalian heart has a limited regenerative capacity. Previous work suggested the heart can regenerate during development and immediately after birth by inducing cardiomyocyte (CM) proliferation; however, this capacity is lost seven days after birth. modRNA gene delivery, the same technology used successfully in the two mRNA vaccines against SARS-CoV-2, can prompt cardiac regeneration, cardiovascular regeneration and cardiac protection. We recently established a novel CM-specific modRNA translational system (SMRTs) that allows modRNA translation only in CMs. We demonstrated that this system delivers potent intracellular genes (e.g., cell cyclepromoting Pkm2), which are beneficial when expressed in one cell type (i.e., CMs) but not others (non-CMs). Here, we identify Lin28a as an important regulator of the CM cell cycle. We show that Lin28a is expressed in CMs during development and immediately after birth, but not during adulthood. We describe that specific delivery of Lin28a into CM, using CM SMRTs, enables CM cell division and proliferation. Further, we determine that this proliferation leads to cardiac repair and better outcome post MI. Moreover, we identify the molecular pathway of Lin28a in CMs. We also demonstrate that Lin28a suppress Let-7 which is vital for CM proliferation, partially due to its suppressive role on cMYC, HMGA2 and K-RAS.
MeSH term(s)	Animals ; Humans ; Adult ; Myocytes, Cardiac ; COVID-19 Vaccines ; Cardiac Surgical Procedures ; Cell Division ; Protein Biosynthesis ; Mammals
Chemical Substances	COVID-19 Vaccines
Language	English
Publishing date	2024-01-30
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	80157-4
ISSN	1095-8584 ; 0022-2828
ISSN (online)	1095-8584
ISSN	0022-2828
DOI	10.1016/j.yjmcc.2024.01.007
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Article: Modified mRNA Formulation and Stability for Cardiac and Skeletal Muscle Delivery.

Żak, Magdalena M / Kaur, Keerat / Yoo, Jimeen / Kurian, Ann Anu / Adjmi, Matthew / Mainkar, Gayatri / Yoon, Seonghun / Zangi, Lior

Pharmaceutics

2023 Volume 15, Issue 9

Abstract: Directly injecting naked or lipid nanoparticle (LNP)-encapsulated modified mRNA (modRNA) allows rapid and efficient protein expression. This non-viral technology has been used successfully in modRNA vaccines against SARS-CoV-2. The main challenges in ... ...

Abstract	Directly injecting naked or lipid nanoparticle (LNP)-encapsulated modified mRNA (modRNA) allows rapid and efficient protein expression. This non-viral technology has been used successfully in modRNA vaccines against SARS-CoV-2. The main challenges in using modRNA vaccines were the initial requirement for an ultra-cold storage to preserve their integrity and concerns regarding unwanted side effects from this new technology. Here, we showed that naked modRNA maintains its integrity when stored up to 7 days at 4 °C, and LNP-encapsulated modRNA for up to 7 days at room temperature. Naked modRNA is predominantly expressed at the site of injection when delivered into cardiac or skeletal muscle. In comparison, LNP-encapsulated modRNA granted superior protein expression but also additional protein expression beyond the cardiac or skeletal muscle injection site. To overcome this challenge, we developed a skeletal-muscle-specific modRNA translation system (skeletal muscle SMRTs) for LNP-encapsulated modRNA. This system allows controlled protein translation predominantly at the site of injection to prevent potentially detrimental leakage and expression in major organs. Our study revealed the potential of the SMRTs platform for controlled expression of mRNA payload delivered intramuscularly. To conclude, our SMRTs platform for LNP-encapsulated modRNA can provide safe, stable, efficient and targeted gene expression at the site of injection.
Language	English
Publishing date	2023-08-22
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2527217-2
ISSN	1999-4923
ISSN	1999-4923
DOI	10.3390/pharmaceutics15092176
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Article ; Online: CCND2 Modified mRNA Activates Cell Cycle of Cardiomyocytes in Hearts With Myocardial Infarction in Mice and Pigs.

Sun, Jiacheng / Wang, Lu / Matthews, Rachel C / Walcott, Gregory P / Lu, Yu-An / Wei, Yuhua / Zhou, Yang / Zangi, Lior / Zhang, Jianyi

Circulation research

2023 Volume 133, Issue 6, Page(s) 484–504

Abstract: Background: Experiments in mammalian models of cardiac injury suggest that the cardiomyocyte-specific overexpression of CCND2 (cyclin D2, in humans) improves recovery from myocardial infarction (MI). The primary objective of this investigation was to ... ...

Abstract	Background: Experiments in mammalian models of cardiac injury suggest that the cardiomyocyte-specific overexpression of CCND2 (cyclin D2, in humans) improves recovery from myocardial infarction (MI). The primary objective of this investigation was to demonstrate that our specific modified mRNA translation system (SMRTs) can induce CCND2 expression in cardiomyocytes and replicate the benefits observed in other studies of cardiomyocyte-specific CCND2 overexpression for myocardial repair. Methods: The CCND2-cardiomyocyte-specific modified mRNA translation system (cardiomyocyte SMRTs) consists of 2 modRNA constructs: one codes for CCND2 and contains a binding site for L7Ae, and the other codes for L7Ae and contains recognition elements for the cardiomyocyte-specific microRNAs miR-1 and miR-208. Thus, L7Ae suppresses CCND2 translation in noncardiomyocytes but is itself suppressed by endogenous miR-1 and -208 in cardiomyocytes, thereby facilitating cardiomyocyte-specific CCND2 expression. Experiments were conducted in both mouse and pig models of MI, and control assessments were performed in animals treated with an SMRTs coding for the cardiomyocyte-specific expression of luciferase or green fluorescent protein (GFP), in animals treated with L7Ae modRNA alone or with the delivery vehicle, and in Sham-operated animals. Results: CCND2 was abundantly expressed in cultured, postmitotic cardiomyocytes 2 days after transfection with the CCND2-cardiomyocyte SMRTs, and the increase was accompanied by the upregulation of markers for cell-cycle activation and proliferation (eg, Ki67 and Aurora B kinase). When the GFP-cardiomyocyte SMRTs were intramyocardially injected into infarcted mouse hearts, the GFP signal was observed in cardiomyocytes but no other cell type. In both MI models, cardiomyocyte proliferation (on day 7 and day 3 after treatment administration in mice and pigs, respectively) was significantly greater, left-ventricular ejection fractions (days 7 and 28 in mice, days 10 and 28 in pigs) were significantly higher, and infarcts (day 28 in both species) were significantly smaller in animals treated with the CCND2-cardiomyocyte SMRTs than in any other group that underwent MI induction. Conclusions: Intramyocardial injections of the CCND2-cardiomyocyte SMRTs promoted cardiomyocyte proliferation, reduced infarct size, and improved cardiac performance in small and large mammalian hearts with MI.
MeSH term(s)	Animals ; Mice ; Cell Cycle ; Cyclin D2/genetics ; Disease Models, Animal ; MicroRNAs/genetics ; MicroRNAs/metabolism ; Myocardial Infarction/metabolism ; Myocytes, Cardiac/metabolism ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Swine
Chemical Substances	Cyclin D2 ; MicroRNAs ; Mirn208 microRNA, mouse ; RNA, Messenger
Language	English
Publishing date	2023-08-11
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	80100-8
ISSN	1524-4571 ; 0009-7330 ; 0931-6876
ISSN (online)	1524-4571
ISSN	0009-7330 ; 0931-6876
DOI	10.1161/CIRCRESAHA.123.322929
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Article ; Online: Synthetic MicroRNAs Stimulate Cardiac Repair.

Zangi, Lior / Hajjar, Roger J

Circulation research

2017 Volume 120, Issue 8, Page(s) 1222–1223

MeSH term(s)	Humans ; MicroRNAs ; Myocardial Infarction ; Myocytes, Cardiac
Chemical Substances	MicroRNAs
Language	English
Publishing date	2017-04-13
Publishing country	United States
Document type	Editorial ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Comment
ZDB-ID	80100-8
ISSN	1524-4571 ; 0009-7330 ; 0931-6876
ISSN (online)	1524-4571
ISSN	0009-7330 ; 0931-6876
DOI	10.1161/CIRCRESAHA.117.310863
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Article ; Online: mRNA-Based Protein Replacement Therapy for the Heart.

Magadum, Ajit / Kaur, Keerat / Zangi, Lior

Molecular therapy : the journal of the American Society of Gene Therapy

2018 Volume 27, Issue 4, Page(s) 785–793

Abstract: Myocardial infarction (MI) and heart failure (HF) are the leading causes of death in the United States and in most other industrialized nations. MI leads to a massive loss of cardiomyocytes (CMs), which are replaced with non-CM cells, leading to scarring ...

Abstract	Myocardial infarction (MI) and heart failure (HF) are the leading causes of death in the United States and in most other industrialized nations. MI leads to a massive loss of cardiomyocytes (CMs), which are replaced with non-CM cells, leading to scarring and, in most cases, HF. The adult mammalian heart has a low intrinsic regenerative capacity, mainly because of cell-cycle arrest in CMs. No effective treatment promoting heart regeneration is currently available. Recent efforts to use DNA-based or viral gene therapy approaches to induce cardiac regeneration post-MI or in HF conditions have encountered major challenges, mostly because of the poor and uncontrolled delivery of the introduced genes. Modified mRNA (modRNA) is a safe, non-immunogenic, efficient, transient, local, and controlled nucleic acid delivery system that can overcome the obstacles to DNA-based or viral approaches for cardiac gene delivery. We here review the use of modRNA in cardiac therapy, to induce cardioprotection and vascular or cardiac regeneration after MI. We discuss the current challenges in modRNA-based cardiac treatment, which will need to be overcome for the application of such treatment to ischemic heart disease.
MeSH term(s)	Animals ; Drug Delivery Systems ; Enzyme Replacement Therapy/methods ; Genetic Therapy/adverse effects ; Genetic Therapy/methods ; Heart Failure/therapy ; Humans ; Myocardial Infarction/therapy ; Myocytes, Cardiac/metabolism ; Nanoparticles ; RNA, Messenger/genetics ; Regeneration ; Transfection
Chemical Substances	RNA, Messenger
Language	English
Publishing date	2018-12-06
Publishing country	United States
Document type	Journal Article ; Review
ZDB-ID	2010592-7
ISSN	1525-0024 ; 1525-0016
ISSN (online)	1525-0024
ISSN	1525-0016
DOI	10.1016/j.ymthe.2018.11.018
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Article ; Online: In Vitro Synthesis of Modified RNA for Cardiac Gene Therapy.

Sultana, Nishat / Sharkar, Mohammad Tofael Kabir / Hadas, Yoav / Chepurko, Elena / Zangi, Lior

Methods in molecular biology (Clifton, N.J.)

2020 Volume 2158, Page(s) 281–294

Abstract: Modified mRNA (modRNA) is a promising new gene therapy approach that has safely and effectively delivered genes into different tissues, including the heart. Current efforts to use DNA-based or viral gene therapy to induce cardiac regeneration ... ...

Abstract	Modified mRNA (modRNA) is a promising new gene therapy approach that has safely and effectively delivered genes into different tissues, including the heart. Current efforts to use DNA-based or viral gene therapy to induce cardiac regeneration postmyocardial infarction (MI) or in heart failure (HF) have encountered key challenges, e.g., genome integration and delayed and noncontrolled expression. By contrast, modRNA is a transient, safe, non-immunogenic, and controlled gene delivery method that is not integrated into the genome. For most therapeutic applications, especially in regenerative medicine, the ability to deliver genes to the heart transiently and with control is vital for achieving therapeutic effect. Additionally, modRNA synthesis is comparatively simple and inexpensive compared to other gene delivery methods (e.g., protein), though a simple, clear in vitro transcription (IVT) protocol for synthesizing modRNA is needed for it to be more widely used. Here, we describe a simple and improved step-by-step IVT protocol to synthesize modRNA for in vitro or in vivo applications.
MeSH term(s)	Animals ; Gene Transfer Techniques ; Genetic Therapy ; Mice ; Myocardial Infarction/genetics ; Myocardial Infarction/therapy ; Myocardium/metabolism ; RNA, Messenger/administration & dosage ; RNA, Messenger/chemistry ; RNA, Messenger/genetics ; Regenerative Medicine ; Transcription, Genetic
Chemical Substances	RNA, Messenger
Language	English
Publishing date	2020-06-29
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-0668-1_21
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Article ; Online: Specific Modified mRNA Translation System.

Magadum, Ajit / Kurian, Ann Anu / Chepurko, Elena / Sassi, Yassine / Hajjar, Roger J / Zangi, Lior

Circulation

2020 Volume 142, Issue 25, Page(s) 2485–2488

MeSH term(s)	Animals ; Cells, Cultured ; Disease Models, Animal ; Gene Transfer Techniques ; Humans ; Interleukin-2 Receptor alpha Subunit/genetics ; Mice ; Mice, Transgenic ; MicroRNAs/genetics ; MicroRNAs/metabolism ; Myocardial Infarction/genetics ; Myocardial Infarction/metabolism ; Myocardial Infarction/pathology ; Myocardial Infarction/therapy ; Myocytes, Cardiac/metabolism ; Myocytes, Cardiac/pathology ; Protein Biosynthesis ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Rats ; Transfection
Chemical Substances	IL2RA protein, human ; Interleukin-2 Receptor alpha Subunit ; MicroRNAs ; Mirn1 microRNA, mouse ; Mirn199 microRNA, mouse ; Mirn208 microRNA, mouse ; RNA, Messenger
Language	English
Publishing date	2020-12-21
Publishing country	United States
Document type	Letter ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	80099-5
ISSN	1524-4539 ; 0009-7322 ; 0069-4193 ; 0065-8499
ISSN (online)	1524-4539
ISSN	0009-7322 ; 0069-4193 ; 0065-8499
DOI	10.1161/CIRCULATIONAHA.120.047211
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