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Article ; Online: Lipid nanoparticles-loaded with toxin mRNA represents a new strategy for the treatment of solid tumors.

Granot-Matok, Yasmin / Ezra, Assaf / Ramishetti, Srinivas / Sharma, Preeti / Naidu, Gonna Somu / Benhar, Itai / Peer, Dan

Theranostics

2023 Volume 13, Issue 11, Page(s) 3497–3508

Abstract: Background and ... ...

Abstract	Background and rationale
MeSH term(s)	Mice ; Animals ; RNA, Messenger/genetics ; Liposomes ; Genetic Therapy ; Nanoparticles ; Neoplasms/therapy ; Bacterial Toxins/genetics
Chemical Substances	Lipid Nanoparticles ; RNA, Messenger ; Liposomes ; Bacterial Toxins
Language	English
Publishing date	2023-06-12
Publishing country	Australia
Document type	Journal Article
ZDB-ID	2592097-2
ISSN	1838-7640 ; 1838-7640
ISSN (online)	1838-7640
ISSN	1838-7640
DOI	10.7150/thno.82228
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Engineering lymphocytes with RNAi.

Ramishetti, Srinivas / Peer, Dan

Advanced drug delivery reviews

2018 Volume 141, Page(s) 55–66

Abstract: Lymphocytes are the gatekeepers of the body's immune system and are involved in pathogenesis if their surveillance is stalled by inhibitory molecules or when they act as mediators for viral entry. Engineering lymphocytes in order to restore their ... ...

Abstract	Lymphocytes are the gatekeepers of the body's immune system and are involved in pathogenesis if their surveillance is stalled by inhibitory molecules or when they act as mediators for viral entry. Engineering lymphocytes in order to restore their functions is an unmet need in immunological disorders, cancer and in lymphotropic viral infections. Recently, the FDA approved several therapeutic antibodies for blocking inhibitory signals on T cells. This has revolutionized the field of solid tumor care, together with chimeric antigen receptor T cell (CAR-T) therapy that did the same for hematological malignancies. RNA interference (RNAi) is a promising approach where gene function can be inhibited in almost all types of cells. However, manipulation of genes in lymphocyte subsets are difficult due to their hard-to-transfect nature and in vivo targeting remains challenging as they are dispersed throughout the body. The ability of RNAi molecules to gain entry into cells is almost impossible without delivery strategy. Nanotechnology approaches are rapidly growing and their impact in the field of drug and gene delivery applications to transport payloads inside cells have been extensively studied. Here we discuss various technologies available for RNAi delivery to lymphocytes. We shed light on the importance of targeting molecules in order to target lymphocytes in vivo. In addition, we discuss recent developments of RNAi delivery to lymphocyte subsets, and detail the potential implication for the future of molecular medicine in leukocytes implicated diseases.
MeSH term(s)	Animals ; Humans ; Lymphocytes/metabolism ; RNA Interference ; RNA, Small Interfering/administration & dosage
Chemical Substances	RNA, Small Interfering
Language	English
Publishing date	2018-12-06
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	639113-8
ISSN	1872-8294 ; 0169-409X
ISSN (online)	1872-8294
ISSN	0169-409X
DOI	10.1016/j.addr.2018.12.002
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Article ; Online: Therapeutic gene silencing of

Chatterjee, Sushmita / Naidu, Gonna Somu / Hazan-Halevy, Inbal / Grobe, Hanna / Ezra, Assaf / Sharma, Preeti / Goldsmith, Meir / Ramishetti, Srinivas / Sprinzak, David / Zaidel-Bar, Ronen / Peer, Dan

Science advances

2023 Volume 9, Issue 14, Page(s) eade4800

Abstract: The potential of microtubule-associated protein targets for cancer therapeutics remains largely unexplored due to the lack of target-specific agents. Here, we explored the therapeutic potential of targeting cytoskeleton-associated protein 5 (CKAP5), an ... ...

Abstract	The potential of microtubule-associated protein targets for cancer therapeutics remains largely unexplored due to the lack of target-specific agents. Here, we explored the therapeutic potential of targeting cytoskeleton-associated protein 5 (CKAP5), an important microtubule-associated protein, with
MeSH term(s)	Humans ; Animals ; Female ; Gene Silencing ; Microtubule-Associated Proteins/metabolism ; RNA, Small Interfering/genetics ; Microtubules/metabolism ; Ovarian Neoplasms/genetics ; Nanoparticles
Chemical Substances	Microtubule-Associated Proteins ; RNA, Small Interfering ; CKAP5 protein, human
Language	English
Publishing date	2023-04-05
Publishing country	United States
Document type	Journal Article
ZDB-ID	2810933-8
ISSN	2375-2548 ; 2375-2548
ISSN (online)	2375-2548
ISSN	2375-2548
DOI	10.1126/sciadv.ade4800
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Article ; Online: Systemic Modulation of Lymphocyte Subsets Using siRNAs Delivered via Targeted Lipid Nanoparticles.

Hazan-Halevy, Inbal / Rosenblum, Daniel / Ramishetti, Srinivas / Peer, Dan

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

2019 Volume 1974, Page(s) 151–159

Abstract: Systemic delivery of RNA interference (RNAi) payloads for manipulation of gene expression in lymphocytes holds a great potential as a novel therapeutic modality for hematological malignancies and autoimmune disorders. However, lymphocytes are among the ... ...

Abstract	Systemic delivery of RNA interference (RNAi) payloads for manipulation of gene expression in lymphocytes holds a great potential as a novel therapeutic modality for hematological malignancies and autoimmune disorders. However, lymphocytes are among the most difficult cells to transfect with RNAi, as they are resistant to conventional transfection reagents and are dispersed throughout the body, making it a challenge to successfully deliver these payloads via systemic administration route. We have developed a strategy to target lymphocytes and deliver RNAi payloads in a cell-specific manner to induce therapeutic gene silencing. This approach utilizes antibodies that decorate lipid nanoparticle surfaces to home into lymphocyte subsets. This approach opens new avenues for discovery of new drug targets and potentially for therapeutics.
MeSH term(s)	Drug Delivery Systems/methods ; Gene Silencing ; Humans ; Lipids/chemistry ; Lipids/genetics ; Lipids/pharmacology ; Lymphocytes/drug effects ; Nanoparticles/chemistry ; RNA, Small Interfering/chemistry ; RNA, Small Interfering/genetics ; RNA, Small Interfering/pharmacology
Chemical Substances	Lipids ; RNA, Small Interfering
Language	English
Publishing date	2019-05-16
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-4939-9220-1_11
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Article ; Online: A Combinatorial Library of Lipid Nanoparticles for Cell Type-Specific mRNA Delivery.

Naidu, Gonna Somu / Yong, Seok-Beom / Ramishetti, Srinivas / Rampado, Riccardo / Sharma, Preeti / Ezra, Assaf / Goldsmith, Meir / Hazan-Halevy, Inbal / Chatterjee, Sushmita / Aitha, Anjaiah / Peer, Dan

Advanced science (Weinheim, Baden-Wurttemberg, Germany)

2023 Volume 10, Issue 19, Page(s) e2301929

Abstract: Ionizable lipid-based nanoparticles (LNPs) are the most advanced non-viral drug delivery systems for RNA therapeutics and vaccines. However, cell type-specific, extrahepatic mRNA delivery is still a major hurdle, hampering the development of novel ... ...

Abstract	Ionizable lipid-based nanoparticles (LNPs) are the most advanced non-viral drug delivery systems for RNA therapeutics and vaccines. However, cell type-specific, extrahepatic mRNA delivery is still a major hurdle, hampering the development of novel therapeutic modalities. Herein, a novel ionizable lipid library is synthesized by modifying hydrophobic tail chains and linkers. Combined with other helper lipids and utilizing a microfluidic mixing approach, stable LNPs are formed. Using Luciferase-mRNA, mCherry mRNA, and Cre mRNA together with a TdTomato animal model, superior lipids forming LNPs for potent cell-type specific mRNA delivery are identified. In vitro assays concluded that combining branched ester tail chains with hydroxylamine linker negatively affects mRNA delivery efficiency. In vivo studies identify Lipid 23 as a liver-trophic, superior mRNA delivery lipid and Lipid 16 as a potent cell type-specific ionizable lipid for the CD11b
MeSH term(s)	Animals ; RNA, Messenger/genetics ; Nanoparticles/chemistry ; Lipids/chemistry
Chemical Substances	Lipid Nanoparticles ; 1-octylnonyl 8-((2-hydroxyethyl)(6-oxo-6-(undecyloxy)hexyl)amino)octanoate (T7OBQ65G2I) ; RNA, Messenger ; Lipids
Language	English
Publishing date	2023-04-24
Publishing country	Germany
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2808093-2
ISSN	2198-3844 ; 2198-3844
ISSN (online)	2198-3844
ISSN	2198-3844
DOI	10.1002/advs.202301929
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Article ; Online: Conformation-sensitive targeting of lipid nanoparticles for RNA therapeutics.

Dammes, Niels / Goldsmith, Meir / Ramishetti, Srinivas / Dearling, Jason L J / Veiga, Nuphar / Packard, Alan B / Peer, Dan

Nature nanotechnology

2021 Volume 16, Issue 9, Page(s) 1030–1038

Abstract: The successful in vivo implementation of gene expression modulation strategies relies on effective, non-immunogenic delivery vehicles. Lipid nanoparticles are one of the most advanced non-viral clinically approved nucleic-acid delivery systems. Yet lipid ...

Abstract	The successful in vivo implementation of gene expression modulation strategies relies on effective, non-immunogenic delivery vehicles. Lipid nanoparticles are one of the most advanced non-viral clinically approved nucleic-acid delivery systems. Yet lipid nanoparticles accumulate naturally in liver cells upon intravenous administration, and hence, there is an urgent need to enhance uptake by other cell types. Here we use a conformation-sensitive targeting strategy to achieve in vivo gene silencing in a selective subset of leukocytes and show potential therapeutic applications in a murine model of colitis. In particular, by targeting the high-affinity conformation of α
MeSH term(s)	Animals ; Colitis/genetics ; Colitis/therapy ; Gene Expression Regulation/drug effects ; Gene Silencing ; Humans ; Integrin alpha4/chemistry ; Integrin alpha4/genetics ; Integrin beta Chains/chemistry ; Integrin beta Chains/genetics ; Lipids/chemistry ; Lipids/pharmacology ; Liver/drug effects ; Mice ; Nanoparticles/chemistry ; Nanoparticles/therapeutic use ; RNA, Small Interfering/genetics ; RNA, Small Interfering/pharmacology
Chemical Substances	Integrin beta Chains ; Lipids ; RNA, Small Interfering ; Integrin alpha4 (143198-26-9)
Language	English
Publishing date	2021-06-17
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2254964-X
ISSN	1748-3395 ; 1748-3387
ISSN (online)	1748-3395
ISSN	1748-3387
DOI	10.1038/s41565-021-00928-x
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Article ; Online: Therapeutic Gene Silencing Using Targeted Lipid Nanoparticles in Metastatic Ovarian Cancer.

Singh, Manu Smriti / Ramishetti, Srinivas / Landesman-Milo, Dalit / Goldsmith, Meir / Chatterjee, Sushmita / Palakuri, Ramesh / Peer, Dan

Small (Weinheim an der Bergstrasse, Germany)

2021 Volume 17, Issue 19, Page(s) e2100287

Abstract: Ovarian cancer is an aggressive tumor owing to its ability to metastasize from stage II onward. Herein, lipid nanoparticles (LNPs) that encapsulate combination of small interfering RNAs (siRNAs), polo-like kinase-1 (PLK1), and eukaryotic translation- ... ...

Abstract	Ovarian cancer is an aggressive tumor owing to its ability to metastasize from stage II onward. Herein, lipid nanoparticles (LNPs) that encapsulate combination of small interfering RNAs (siRNAs), polo-like kinase-1 (PLK1), and eukaryotic translation-initiation factor 3c (eIF3c), to target different cellular pathways essential for ovarian cancer progression are generated. The LNPs are further modified with hyaluronan (tNPs) to target cluster of differentiation 44 (CD44) expressing cells. Interestingly, hyaluronan-coated LNPs (tNPs) prolong functional activity and reduce growth kinetics of spheroids in in vitro assay as compared to uncoated LNPs (uNPs) due to ≈1500-fold higher expression of CD44. Treatment of 2D and 3D cultured ovarian cancer cells with LNPs encapsulating both siRNAs result in 85% cell death and robust target gene silencing. In advanced orthotopic ovarian cancer model, intraperitoneal administration of LNPs demonstrates CD44 specific tumor targeting of tNPs compared to uNPs and robust gene silencing in tissues involved in ovarian cancer pathophysiology. At very low siRNA dose, enhanced overall survival of 60% for tNPs treated mice is observed compared to 10% and 20% for single siRNA-, eIF3c-tNP, and PLK1-tNP treatment groups, respectively. Overall, LNPs represent promising platform in the treatment of advanced ovarian cancer by improving median- and overall-survival.
MeSH term(s)	Animals ; Female ; Gene Silencing ; Humans ; Lipids ; Mice ; Nanoparticles ; Ovarian Neoplasms ; RNA, Small Interfering
Chemical Substances	Lipids ; RNA, Small Interfering
Language	English
Publishing date	2021-04-07
Publishing country	Germany
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2168935-0
ISSN	1613-6829 ; 1613-6810
ISSN (online)	1613-6829
ISSN	1613-6810
DOI	10.1002/smll.202100287
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Article ; Online: A Combinatorial Library of Lipid Nanoparticles for Cell Type‐Specific mRNA Delivery

Gonna Somu Naidu / Seok‐Beom Yong / Srinivas Ramishetti / Riccardo Rampado / Preeti Sharma / Assaf Ezra / Meir Goldsmith / Inbal Hazan‐Halevy / Sushmita Chatterjee / Anjaiah Aitha / Dan Peer

Advanced Science, Vol 10, Iss 19, Pp n/a-n/a (2023)

2023

Abstract: Abstract Ionizable lipid‐based nanoparticles (LNPs) are the most advanced non‐viral drug delivery systems for RNA therapeutics and vaccines. However, cell type‐specific, extrahepatic mRNA delivery is still a major hurdle, hampering the development of ... ...

Abstract	Abstract Ionizable lipid‐based nanoparticles (LNPs) are the most advanced non‐viral drug delivery systems for RNA therapeutics and vaccines. However, cell type‐specific, extrahepatic mRNA delivery is still a major hurdle, hampering the development of novel therapeutic modalities. Herein, a novel ionizable lipid library is synthesized by modifying hydrophobic tail chains and linkers. Combined with other helper lipids and utilizing a microfluidic mixing approach, stable LNPs are formed. Using Luciferase‐mRNA, mCherry mRNA, and Cre mRNA together with a TdTomato animal model, superior lipids forming LNPs for potent cell‐type specific mRNA delivery are identified. In vitro assays concluded that combining branched ester tail chains with hydroxylamine linker negatively affects mRNA delivery efficiency. In vivo studies identify Lipid 23 as a liver‐trophic, superior mRNA delivery lipid and Lipid 16 as a potent cell type‐specific ionizable lipid for the CD11bhi macrophage population without an additional targeting moiety. Finally, in vivo mRNA delivery efficiency and toxicity of these LNPs are compared with SM‐102‐based LNP (Moderna's LNP formulation) and are shown to be cell‐specific compared to SM‐102‐based LNPs. Overall, this study suggests that a structural combination of tail and linker can drive a novel functionality of LNPs in vivo.
Keywords	cell type‐specific mRNA delivery ; combinatorial lipid nanoparticles ; mRNA delivery ; Science ; Q
Subject code	610
Language	English
Publishing date	2023-07-01T00:00:00Z
Publisher	Wiley
Document type	Article ; Online
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Article ; Online: Advances in RNAi therapeutic delivery to leukocytes using lipid nanoparticles.

Ramishetti, Srinivas / Landesman-Milo, Dalit / Peer, Dan

Journal of drug targeting

2016 Volume 24, Issue 9, Page(s) 780–786

Abstract: Small interfering RNAs (siRNAs) therapeutics has advanced into clinical trials for liver diseases and solid tumors, but remain a challenge for manipulating leukocytes fate due to lack of specificity and safety issues. Leukocytes ingest pathogens and ... ...

Abstract	Small interfering RNAs (siRNAs) therapeutics has advanced into clinical trials for liver diseases and solid tumors, but remain a challenge for manipulating leukocytes fate due to lack of specificity and safety issues. Leukocytes ingest pathogens and defend the body through a complex network. They are also involved in the pathogeneses of inflammation, viral infection, autoimmunity and cancers. Modulating gene expression in leukocytes using siRNAs holds great promise to treat leukocyte-mediated diseases. Leukocytes are notoriously hard to transduce with siRNAs and are spread throughout the body often located deep in tissues, therefore developing an efficient systemic delivery strategy is still a challenge. Here, we discuss recent advances in siRNA delivery to leukocyte subsets such as macrophages, monocytes, dendritic cells and lymphocytes. We focus mainly on lipid-based nanoparticles (LNPs) comprised of new generation of ionizable lipids and their ability to deliver siRNA to primary or malignant leukocytes in a targeted manner. Special emphasis is made on LNPs targeted to subsets of leukocytes and we detail a novel microfluidic mixing technology that could aid in changing the landscape of process development of LNPs from a lab tool to a potential novel therapeutic modality.
MeSH term(s)	Gene Transfer Techniques ; Genetic Therapy/methods ; Lipids/chemistry ; Nanoparticles/chemistry ; RNA Interference
Chemical Substances	Lipids
Language	English
Publishing date	2016-05-04
Publishing country	England
Document type	Journal Article ; Review ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	1187110-6
ISSN	1029-2330 ; 1061-186X
ISSN (online)	1029-2330
ISSN	1061-186X
DOI	10.3109/1061186X.2016.1172587
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Article ; Online: Dual-Targeted Lipid Nanotherapeutic Boost for Chemo-Immunotherapy of Cancer.

Yong, Seok-Beom / Ramishetti, Srinivas / Goldsmith, Meir / Diesendruck, Yael / Hazan-Halevy, Inbal / Chatterjee, Sushmita / Somu Naidu, Gonna / Ezra, Assaf / Peer, Dan

Advanced materials (Deerfield Beach, Fla.)

2022 Volume 34, Issue 13, Page(s) e2106350

Abstract: Chemo-immunotherapy is a combination of "standard-of-care" chemotherapy with immunotherapy and it is considered the most advanced therapeutic modality for various types of cancers. However, many cancer patients still poorly respond to current regimen of ... ...

Abstract	Chemo-immunotherapy is a combination of "standard-of-care" chemotherapy with immunotherapy and it is considered the most advanced therapeutic modality for various types of cancers. However, many cancer patients still poorly respond to current regimen of chemo-immunotherapy and suggest nanotherapeutics as a boosting agent. Recently, heme oxygenase-1 (HO1) is shown to act as an immunotherapeutic molecule in tumor myeloid cells, in addition to general chemoresistance function in cancer cells suggesting that HO1-targeted therapeutics can become a novel, optimal strategy for boosting chemo-immunotherapy in the clinic. Currently the available HO1-inhibitors demonstrate serious adverse effects in clinical use. Herein, tumor myeloid cell- and cancer cell-dual targeted HO1-inhibiting lipid nanotherapeutic boost (T-iLNTB) is developed using RNAi-loaded lipid nanoparticles. T-iLNTB-mediated HO1-inhibition sensitizes cancer cells to "standard-of-care" chemotherapeutics by increasing immunogenic cell death, and directly reprograms tumor myeloid cells with distinguished phenotype. Furthermore, tumor myeloid cell reprogramming by T-iLNTB induces CD8
MeSH term(s)	Humans ; Immunotherapy ; Lipids ; Liposomes ; Nanoparticles ; Neoplasms/drug therapy ; Tumor Microenvironment
Chemical Substances	Lipid Nanoparticles ; Lipids ; Liposomes
Language	English
Publishing date	2022-02-10
Publishing country	Germany
Document type	Journal Article
ZDB-ID	1474949-X
ISSN	1521-4095 ; 0935-9648
ISSN (online)	1521-4095
ISSN	0935-9648
DOI	10.1002/adma.202106350
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