Genetic engineering of mesenchymal stem cells for regenerative medicineby Adam Nowakowski, Piotr Walczak, Miroslaw Janowski, Barbara Lukomska

Stem Cells and Development

About

Year
2015
DOI
10.1089/scd.2015.0062
Subject
Developmental Biology / Cell Biology / Hematology

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Genetic engineering of mesenchymal stem cells for regenerative medicine

Adam Nowakowski 1 , Piotr Walczak 2-4 , Miroslaw Janowski 1-3 , Barbara Lukomska 1

Affiliations: 1

NeuroRepair Department, Mossakowski Medical Research Centre, Polish Academy of Sciences,

Warsaw, Poland 2

Russell H. Morgan Dept. of Radiology and Radiological Science, Division of MR Research, 3

Cellular Imaging Section and Vascular Biology Program, Institute for Cell Engineering, The Johns

Hopkins University School of Medicine, Baltimore, USA 4

Dept of Radiology, Faculty of Medical Sciences, University of Warmia and Mazury, Olsztyn, Poland

Running Head: Genetic engineering of mesenchymal stem cells for regenerative medicine

Keywords: mesenchymal stem cells, transfection, gene therapy

Corresponding: Dr. Adam Nowakowski anowakowski@imdik.pan.pl

Abstract

Mesenchymal stem cells (MSCs), which can be obtained from various organs and easily propagated in vitro, are one of the most extensively used types of stem cells and have been shown to be efficacious in a broad set of diseases. The unique and highly desirable properties of MSCs include high migratory capacities toward injured areas, immunomodulatory features, and the natural ability to differentiate into connective tissue phenotypes. These phenotypes include bone and cartilage, and these properties predispose MSCs to be therapeutically useful. In addition, MSCs elicit their therapeutic effects by paracrine actions, in which the metabolism of target tissues is modulated. Genetic engineering methods can greatly amplify these properties and broaden the therapeutic capabilities of MSCs, including transdifferentiation toward diverse cell lineages. However, cell engineering can also affect

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D ev el op m en t

G en et ic e ng in ee rin g of m es en ch ym al st em c el ls fo r r eg en er at iv e m ed ic in e (do i: 1 0.1 08 9/s cd .20 15 .00 62 )

Th is ar tic le h as b ee n pe er -re vi ew ed a nd a cc ep te d fo r p ub lic at io n, b ut h as y et to u nd er go c op ye di tin g an d pr oo f c or re ct io n.

T he fi na l p ub lis he d ve rs io n m ay d iff er fr om th is pr oo f. 2 2 safety and increase the cost of therapy based on MSCs; thus, the advantages and disadvantages of these procedures should be discussed. In this review, the latest applications of genetic engineering methods for MSCs with regenerative medicine purposes are presented.

Introduction

Since their first description in the 1960s, mesenchymal stem cells (MSCs) have been of interest to researchers who use their therapeutic properties for the treatment of various pathological conditions. Regenerative medicine and oncology are the major fields of potential application [1-3].

Considering the wide breadth of literature on MSC engineering, we chose to limit our review to regenerative medicine. The simplicity of obtaining these cells from different organs and the subsequent ease of propagation in vitro [4], as well as the potential for autologous transplantation, makes them a tempting possibility for clinical use. However, special emphasis must be placed on the correct characterization of the obtained cells from different sources and species, to be able to properly compare and make right conclusions on the ground of multiples studies [5,6]. Independently of their original tissue location, MSCs have a natural ability to differentiate into mature mesenchymal phenotypes and form bone or cartilage, so they can be used in the treatment of injuries where these types of tissues are in need of repair [7,8]. What is more, MSCs act through paracrine effects, releasing a plethora of beneficial compounds [9]. While Prochymal (Mesoblast Ltd.), a preparation of allogeneic

MSCs, has recently received conditional approval from the FDA, the engineered MSCs did not reach the level of clinical trials. However, genetically engineered neural stem cells are being tested in clinical trial sponsored by ReNeuron (NCT01151124 and NCT02117635, Clinicaltrials.gov).

However, the diverse biological improvements offered by genetic engineering have the potential to greatly increase the therapeutically useful qualities of MSCs, and tailor them to specific diseases, so they are more probably to be used clinically in the future. A variety of studies have investigated the engineering of MSCs, aiming at stimulation of their direct differentiation toward endothelial cells and angiogenesis [10-16]. In addition, in in vitro (Fig. 1) culture conditions and in vivo studies (Fig. 2),

MSCs can be differentiated into lineage-specific cells or lineage-specific-like cells, such as

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G en et ic e ng in ee rin g of m es en ch ym al st em c el ls fo r r eg en er at iv e m ed ic in e (do i: 1 0.1 08 9/s cd .20 15 .00 62 )

Th is ar tic le h as b ee n pe er -re vi ew ed a nd a cc ep te d fo r p ub lic at io n, b ut h as y et to u nd er go c op ye di tin g an d pr oo f c or re ct io n.

T he fi na l p ub lis he d ve rs io n m ay d iff er fr om th is pr oo f. 3 3 hepatocytes [17], cardiomyocytes [18], pacemaking cells [19], and neuronal cells [20]. Genetically modified MSCs can also be used to ameliorate several neurological disorders by exploiting their paracrine characteristics [21,22]. Moreover, MSCs could be modified to gain anti-fibrotic properties [23]. Finally, MSCs can be committed to overproduce anti-inflammatory cytokines that alleviate local tissue inflammatory states [24]. The abundance of genetic engineering modifications in MSCs quoted in this review was summarised in Tab.1.

This review focuses on a set of examples of the various genetic modifications of MSCs in order to enhance and/or expand MSCs differentiation potential toward multiple cell types for therapeutic purposes, which have been published in recent years, revealing the growing interest directed at these cells and the diversity of strategies (Fig. 3).

Vectors, methods, and approaches to MSC engineering

There are various vectors used for delivery of genetic materials to the cells, such as viruses [25], DNA plasmids [26,27], constructs of minimalistic, immunologically defined gene expression (MIDGE) [28], transposons constructs such as the Sleeping Beauty System [29], mRNA [30], miRNA [31], and siRNA [32]. While viruses directly transduce cells, non-viral methods require specific methods to cross the cell membrane, such as lipofection or electroporation. The vectors and methods for crossing the cell membrane by genetic material have been recently extensively reviewed [33].