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Sonodelivery in Skeletal Muscle: Current Approaches and Future Potential

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Sonodelivery in Skeletal Muscle: Current Approaches and Future Potential bioengineering Review Sonodelivery in Skeletal Muscle: Current Approaches and Future Potential Richard E. Decker 1 , Zachary E. Lamantia 1, Todd S. Emrick 2 and Marxa L. Figueiredo 1,* 1 Department of Basic Medical Sciences, Purdue University, 625 Harrison St., West Lafayette, IN 47907, USA; [email protected] (R.E.D.); [email protected] (Z.E.L.) 2 Department of Polymer Science & Engineering, University of Massachusetts, 120 Governors Drive, Amherst, MA 01003, USA; [email protected] * Correspondence: mlfi[email protected]; Tel.: +1-765-494-5790 Received: 17 July 2020; Accepted: 4 September 2020; Published: 9 September 2020 Abstract: There are currently multiple approaches to facilitate gene therapy via intramuscular gene delivery, such as electroporation, viral delivery, or direct DNA injection with or without polymeric carriers. Each of these methods has benefits, but each method also has shortcomings preventing it from being established as the ideal technique. A promising method, ultrasound-mediated gene delivery (or sonodelivery) is inexpensive, widely available, reusable, minimally invasive, and safe. Hurdles to utilizing sonodelivery include choosing from a large variety of conditions, which are often dependent on the equipment and/or research group, and moderate transfection efficiencies when compared to some other gene delivery methods. In this review, we provide a comprehensive look at the breadth of sonodelivery techniques for intramuscular gene delivery and suggest future directions for this continuously evolving field. Keywords: sonoporation; sonodelivery; ultrasound; intramuscular gene delivery; gene therapy 1. Introduction Gene therapy shows increasing promise for the treatment of a wide range of diseases, from cancer to musculoskeletal and immunological disorders [1]. Although the basic premise of gene therapy is relatively simple on the surface—to introduce a gene or genes into a patient’s cells/tissue that will allow the body to produce a therapeutic protein or facilitate a genetic modification—every step of the process holds its own challenges. One of the major challenges associated with making gene therapy widely (and safely) accessible is that of gene delivery in vivo. There are currently many methods for delivering DNA in vitro at high efficiencies, including electroporation, lipid-based transfection, and viral delivery. Each of these methods presents potential challenges for clinical translation, including the cost and time of production, the invasiveness of the technique, the efficiency of delivery, and/or safety concerns related to an immune response or unintended genomic integration of the therapeutic gene. The development of a gene delivery technique that is inexpensive, widely available, reusable, minimally invasive, and safe will catalyze progress in gene therapy. One promising technique that meets all of these criteria has been under investigation for many years but has not yet reached widespread use: ultrasound-mediated or sonoporation gene delivery, herein referred to as sonodelivery [2,3]. As its name suggests, this approach involves using ultrasound to deliver non-viral vectors encoding therapeutic proteins into skeletal muscle, allowing the body to then produce its own therapeutic. Despite the work that has been done to explore the practicality of ultrasound-mediated intramuscular gene delivery, there is, to our knowledge, no comprehensive review of the most common approaches and technical conditions used for this promising technology. This review aims to fill that void and to discuss the future work needed for and the potential of establishing sonodelivery as a key player in the gene therapy field. Bioengineering 2020, 7, 107; doi:10.3390/bioengineering7030107 www.mdpi.com/journal/bioengineering Bioengineering 2020, 7, 107 2 of 16 Bioengineering 2020, 7, x FOR PEER REVIEW 2 of 16 2. Methods for Cellular and Intramuscular Gene Delivery 2. Methods for Cellular and Intramuscular Gene Delivery There are many established methodologies for delivering plasmid DNA (pDNA) to muscles, each withThere its are own many advantages established and methodologies disadvantages for (Figure delivering1). To highlightplasmid DNA these (pDNA) and provide to muscles, context foreach the with importance its own advantages of sonodelivery, and disadvantages we will provide (Figure a brief 1). background To highlight on these four and of the provide most commoncontext pDNAfor thedelivery importance approaches of sonodelivery, (excluding we sonodelivery).will provide a brief background on four of the most common pDNA delivery approaches (excluding sonodelivery). Figure 1. Common approaches for delivery of plasmid DNA into skeletal muscle and their associated advantagesFigure 1. Common (green) andapproaches disadvantages for delivery (red). of plasmid DNA into skeletal muscle and their associated advantages (green) and disadvantages (red). 2.1. Direct Injection of Naked DNA 2.1. Direct Injection of Naked DNA The oldest and most basic method of intramuscular DNA delivery is injection of naked DNA directlyThe into oldest the targetand most muscle. basic This method method of intramus yields acular low immunogenicDNA delivery response is injectionin vivoof nakedand isDNA both widelydirectly available into the andtarget inexpensive muscle. This [4]. method Plasmids yields are low-costa low immunogenic and low maintenance response in and vivo intramuscular and is both injectionswidely available are straightforward and inexpensive (though [4]. notPlasmids necessarily are low-cost easy) to and administer, low maintenance making this and method intramuscular one of the simplestinjections to are apply. straightforward However, it su (thoughffers from not a verynecessarily low effi easy)ciency to [5 administer,], which has making prevented this direct method injection one ofof naked the simplest DNA from to apply. becoming However, a viable it suffers technique. from a It very has beenlow efficiency reported that[5], which swelling has the prevented muscles direct before injectioninjection (e.g., of naked pretreating DNA from with becoming a hypertonic a viable solution) technique. improves It has expression been reported of proteins that forswelling which the the transfectedmuscles before genes injection code [4 (e.g.,,6,7]. pretreating Similarly, introducing with a hypertonic myotoxic solution) agents improves to the target expression muscle of a proteins few days priorfor which to DNA the injection transfected has beengenes shown code [4,6,7]. to improve Simila proteinrly, introducing expression, myotoxic but it is notagents yet clearto the whether target thismuscle improvement a few days is prior due to to DNA improved injection uptake has ofbeen naked shown DNA to improve or improved protein gene expression, expression but following it is not uptakeyet clear [4 ,whether8]. With this all intramuscularimprovement is gene due delivery,to improved injection uptake technique of naked isDNA crucial or improved [9]. A properly gene localizedexpression injection following at optimal uptake depth[4,8]. With is essential, all intramuscular but is increasingly gene delivery, challenging injection as the technique size of the is crucialin vivo model[9]. A properly increases localized [4]. Injections injection are at therefore optimal depth difficult is essential, to standardize, but is increasingly as optimal techniqueschallenging requireas the trainingsize of the and in extensive vivo model practice. increases One [4]. method Injections thathas are beentherefore used difficult to increase to standardize, injection accuracy as optimal when deliveringtechniques a varietyrequire oftraining payloads and is theextensive utilization practice of imaging. One method ultrasound that tohas guide been the used injection to increase [10–13 ], althoughinjection thisaccuracy too requires when delivering expertise a andvariety extensive of payloads practice. is theAnother utilization complication of imagingassociated ultrasound with to increasedguide the test injection animal [10–13], size is that although of decreased this too gene requires uptake expertise/expression. and Thisextensive may bepractice. due to increasedAnother levelscomplication of connective associated tissue with in increased larger animals, test animal which size either is that acts of todecreased disperse gene the nakeduptake/expression. DNA or as a barrierThis may to the be DNA, due to preventing increased it levels from enteringof connective muscle tissue cells [14in]. largerDespite animals, extensive whichresearch either involving acts to disperse the naked DNA or as a barrier to the DNA, preventing it from entering muscle cells [14]. direct injection of naked DNA, DNA uptake and protein expression levels remain low. Thus, efforts Despite extensive research involving direct injection of naked DNA, DNA uptake and protein to improve the efficiency of DNA delivery, including both biochemical and physical approaches, expression levels remain low. Thus, efforts to improve the efficiency of DNA delivery, including both have led to strategies utilizing polymer and nanoparticle coupling, viral vectors, and electroporation, biochemical and physical approaches, have led to strategies utilizing polymer and nanoparticle among others. coupling, viral vectors, and electroporation, among others. Bioengineering 2020, 7, 107 3 of 16 2.2. Polymers and Nanoparticles Polymer and/or nanoparticle conjugation to DNA involves incubating DNA with the polymer/nanoparticle sample prior to delivery to
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