Exosome-mediated Delivery of Hydrophobically Modified siRNA for Huntingtin mRNA Silencing

Didiot, Marie-Cécile; Hall, Lauren M; Coles, Andrew H.; Haraszti, Reka A.; Godinho, Bruno M.D.C.; Chase, Kathryn; Sapp, Ellen; Ly, Socheata; Alterman, Julia F.; Hassler, Matthew R.; Echeverria, Dimas; Raj, Lakshmi; Morrissey, David; DiFiglia, Marian; Aronin, Neil; Khvorova, Anastasia

doi:10.1038/mt.2016.126

Cited by 365 publications

(289 citation statements)

References 49 publications

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“…4,[18][19][20] The intrinsic cell-targeting property of exosomes can be further enhanced by using genetic engineering techniques to introduce specific proteins to their surface, including ligands for receptors (Apo-A1) or antibodies directed against tumor biomarkers. 21,22 Alvarez-Erviti et al, for example, engineered exosomes produced by dendritic cells to express the neuron-specific rabies viral glycoprotein peptide, which binds to the acetylcholine receptor expressed on neuronal cells. These exosomes were shown to cross the BBB.…”

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confidence: 99%

Engineered exosome-mediated delivery of functionally active miR-26a and its enhanced suppression effect in HepG2 cells

Liang

Kan

Zhu

et al. 2018

IJN

206

131

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Introduction Exosomes are closed-membrane nanovesicles that are secreted by a variety of cells and exist in most body fluids. Recent studies have demonstrated the potential of exosomes as natural vehicles that target delivery of functional small RNA and chemotherapeutics to diseased cells. Methods In this study, we introduce a new approach for the targeted delivery of exosomes loaded with functional miR-26a to scavenger receptor class B type 1-expressing liver cancer cells. The tumor cell-targeting function of these engineered exosomes was introduced by expressing in 293T cell hosts, the gene fusion between the transmembrane protein of CD63 and a sequence from Apo-A1. The exosomes harvested from these 293T cells were loaded with miR-26a via electroporation. Results The engineered exosomes were shown to bind selectively to HepG2 cells via the scavenger receptor class B type 1–Apo-A1 complex and then internalized by receptor-mediated endocytosis. The release of miR-26a in exosome-treated HepG2 cells upregulated miR-26a expression and decreased the rates of cell migration and proliferation. We also presented evidence that suggest cell growth was inhibited by miR-26a-mediated decreases in the amounts of key proteins that regulate the cell cycle. Conclusion Our gene delivery strategy can be adapted to treat a broad spectrum of cancers by expressing proteins on the surface of miRNA-loaded exosomes that recognize specific biomarkers on the tumor cell.

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mentioning

confidence: 99%

Engineered exosome-mediated delivery of functionally active miR-26a and its enhanced suppression effect in HepG2 cells

Liang

Kan

Zhu

et al. 2018

IJN

206

131

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“…The therapeutics that primarily could benefit from EV delivery are RNA therapeutics as delivery of RNA molecules is restricted due to its hydrophilicity and negative charges (37). Since, EVs naturally contain and are shown to transfer this RNA cargo to the recipient cells via cell-cell communication, delivery through EVs could theoretically achieve the desired phenotypic and gene expression changes in the target cells, as has been previously demonstrated (38–40). …”

Section: Evs As Therapeuticsmentioning

confidence: 84%

“…Didiot, et al, explored the use of cholesterol modified huntingtin gene (Htt) siRNA to improve loading into EVs (40). A chemically modified Htt siRNA with a phosphorothioated tail, and 2′-fluoro or 2′-O-methyl pyrimidine modifications were used to protect against nuclease degradation.…”

Section: Loading Mechanismsmentioning

confidence: 99%

“…They showed that even though the cholesterol siRNA was loaded effectively onto the EVs, and were taken up by dendritic JAWS11 and lung epithelial B16F10 cells, functional siRNA could not be delivered in vitro. The cause of the inconsistency in efficacy between these two studies is unclear (40, 80). It may be due to the type of EV and cells used since both studies used different EV and recipient cell combinations.…”

Section: Loading Mechanismsmentioning

confidence: 99%

“…It may be due to the type of EV and cells used since both studies used different EV and recipient cell combinations. In short, cholesterol conjugation is a straightforward method to load therapeutic oligonucleotides into EVs (40). …”

Section: Loading Mechanismsmentioning

confidence: 99%

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Achieving the Promise of Therapeutic Extracellular Vesicles: The Devil is in Details of Therapeutic Loading

et al. 2017

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Extracellular vesicles (EVs) represent a class of cell secreted organelles which naturally contain biomolecular cargo such as miRNA, mRNA and proteins. EVs mediate intercellular communication, enabling the transfer of functional nucleic acids from the cell of origin to the recipient cells. In addition, EVs make an attractive delivery vehicle for therapeutics owing to their increased stability in circulation, biocompatibility, low immunogenicity and toxicity profiles. EVs can also be engineered to display targeting moieties on their surfaces which enables targeting to desired tissues, organs or cells. While much has been learned on the role of EVs as cell communicators, the field of therapeutic EV application is currently under development. Critical to the future success of EV delivery system is the description of methods by which therapeutics can be successfully and efficiently loaded within the EVs. Two methods of loading of EVs with therapeutic cargo exist, endogenous and exogenous loading. We have therefore focused this review on describing the various published approaches for loading EVs with therapeutics.

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Emerging Prospects of Exosomes for Cancer Treatment: From Conventional Therapy to Immunotherapy

et al. 2020

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remains unclear how and why cancer occurs and progresses. Using an evolutionary approach, the concept of a complex adaptive system (CAS) was developed to describe the behaviors of cancer tumorigenesis, [2] which may suggest a frameshift away from the limitations of current approaches that mainly address the importance of alteration in specific target molecules in cancer cells. This change in perspectives toward tumors, not simply as a disease to be cured but also as CAS, is expected to be the cornerstone of the paradigm shift toward innovative cancer therapies. [3] The human immune system can also be viewed as a CAS and therefore expected to initiate self-defense mechanisms against cancer in a complex adaptive manner as long as it recognizes the cancer cells as the non-self-signals. Consequently, the key to controlling cancer could lie in understanding how to manipulate the immune system and strengthen its defenses against cancer. The concept of facilitating the immune system to fight against cancer was first suggested in the late 1800s by Dr. Wiliam Coley, who was the first to observe anti-tumor effects after intratumoral injection of microbe-derived toxins. [4] Since then, the field of cancer immunotherapy research has flourished, resulting in clinical achievements such as immune checkpoint blockades and chimeric antigen receptor T cell (CAR-T) therapy. [5] However, immune suppression resistance mechanisms have simultaneously been identified that have impeded favorable response to cancer immunotherapy. [6,7] Exosome-based cancer therapies have emerged as a potential option for overcoming these limitations to the effects of current cancer therapies due to their pathophysiological efficacy against tumors. [8] Exosomes are secreted externally by cells and are found ubiquitously in blood, urine, saliva, cerebrospinal fluid, pleural fluid, and breast milk. [9,10] The distinction between different types of extracellular vesicles (EVs) is unclear; however, they are conventionally classified as either ectosomes (microvesicles or microparticles) or exosomes. [11] While ectosomes are formed by the outward budding of the plasma membrane, exosomes are formed from multivesicular bodies (MVB) containing intraluminal vesicles via inward budding of the late endosome, which later fuses to the membrane. The formed vesicles are then secreted via a process known as exocytosis (Figure 1). The two types of vesicle also differ in diameter, Exosomes are a class of extracellular vesicles of around 100 nm in diameter that are secreted by most cells and contain various bioactive molecules reflecting their cellular origin and mediate intercellular communication. Studies of these exosomal features in tumor pathogenesis have led to the development of therapeutic and diagnostic approaches using exosomes for cancer therapy. Exosomes have many advantages for conveying therapeutic agents such as small interfering RNAs, microRNAs, membrane-associated proteins, and chemotherapeutic compounds; thus, they are considered a prime candidate as a deliv...

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Exosome-mediated Delivery of Hydrophobically Modified siRNA for Huntingtin mRNA Silencing

Cited by 365 publications

References 49 publications

Engineered exosome-mediated delivery of functionally active miR-26a and its enhanced suppression effect in HepG2 cells

Engineered exosome-mediated delivery of functionally active miR-26a and its enhanced suppression effect in HepG2 cells

Achieving the Promise of Therapeutic Extracellular Vesicles: The Devil is in Details of Therapeutic Loading

Emerging Prospects of Exosomes for Cancer Treatment: From Conventional Therapy to Immunotherapy

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