Nonviral Transfection With Brachyury Reprograms Human Intervertebral Disc Cells to a Pro‐Anabolic Anti‐Catabolic/Inflammatory Phenotype: A Proof of Concept Study

Tang, Shirley; Richards, Justin; Khan, Safdar N.; Hoyland, Judith A.; Gallego‐Perez, Daniel; Higuita‐Castro, Natalia; Walter, Benjamin A.; Purmessur, Devina

doi:10.1002/jor.24408

Cited by 24 publications

(49 citation statements)

References 51 publications

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“…From early organogenesis, its expression is restricted to axial mesoderm/notochord lineage and further maintained after birth in NC constituting the NP, the core of the IVD, in mouse and in human [64,65]. In a recent report, T-encoding plasmid transfection was shown to reprogram mildly degenerate human CLC in vitro to a healthy NP-like phenotype with increased expression of key NP markers and significant proteoglycan/glycosaminoglycan accumulation [66]. Another interesting study also used T-encoding plasmid transfection to differentiate hiPSCs toward an NLC phenotype capable of synthesizing a proteoglycan-rich matrix and playing a protective role in the catabolic environment of injury-induced porcine disc model [67].…”

Section: Discussionmentioning

confidence: 99%

NOTO Transcription Factor Directs Human Induced Pluripotent Stem Cell-Derived Mesendoderm Progenitors to a Notochordal Fate

Colombier

Halgand

Chédeville

et al. 2020

Cells

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The founder cells of the Nucleus pulposus, the centre of the intervertebral disc, originate in the embryonic notochord. After birth, mature notochordal cells (NC) are identified as key regulators of disc homeostasis. Better understanding of their biology has great potential in delaying the onset of disc degeneration or as a regenerative-cell source for disc repair. Using human pluripotent stem cells, we developed a two-step method to generate a stable NC-like population with a distinct molecular signature. Time-course analysis of lineage-specific markers shows that WNT pathway activation and transfection of the notochord-related transcription factor NOTO are sufficient to induce high levels of mesendoderm progenitors and favour their commitment toward the notochordal lineage instead of paraxial and lateral mesodermal or endodermal lineages. This study results in the identification of NOTO-regulated genes including some that are found expressed in human healthy disc tissue and highlights NOTO function in coordinating the gene network to human notochord differentiation.

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Section: Discussionmentioning

confidence: 99%

NOTO Transcription Factor Directs Human Induced Pluripotent Stem Cell-Derived Mesendoderm Progenitors to a Notochordal Fate

Colombier

Halgand

Chédeville

et al. 2020

Cells

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“…The effect of transfecting GDF6 was examined using this protocol and system; however, due to potential limitations with the specific GDF6 plasmid used, no increase in ECM proteins could be observed. Tang et al used this same Neon transfection system to examine the effect of electroporating developmental transcription factor Brachyury into human nucleus pulposus cells from cadavers and patients undergoing surgery for low back pain in 3D in vitro culture (Tang S. et al, 2019). In this study, significant increases in Brachyury were observed up to 4 weeks, together with improvements in IVD phenotypic markers FOXF1, KRT19, and SOX9 and decreases in inflammatory/catabolic/pain markers IL1−β, IL-6, NGF, and MMP-13 compared to transfected sham vector control cells.…”

Section: Physical Gene Vector Methods For the Intervertebral Diskmentioning

confidence: 99%

“…Electroporation (Gehl, 2003;Wells, 2004;Glover et al, 2005;Mehier-Humbert and Guy, 2005;Liu et al, 2006;Al-Dosari and Gao, 2009;Boukany et al, 2011;Guo and Huang, 2012;Mellott et al, 2013;Song et al, 2015;Tschon et al, 2016;Tsuchiya et al, 2017;Vroomen et al, 2017;Kawai et al, 2018;Melancon et al, 2018;Shapiro et al, 2018;Shi B. et al, 2018;Tang S. et al, 2019;Bono et al, 2020) Rapid and straightforward transfection method.…”

Section: Advantages Limitations Applicationsmentioning

confidence: 99%

“…Spinal cord and spinal nerves (Tschon et al, 2016) Periodontal tissue (Kawai et al, 2018) Tibial tumor (Melancon et al, 2018) Tumors in liver, lung, and kidney (Vroomen et al, 2017) Bone -Femur (Song et al, 2015) Periodontal ligament (Tsuchiya et al, 2017) IVD (Bucher et al, 2013;May et al, 2017;Tang S. et al, 2019) Skin (Jafari et al, 2018;Pasquet et al, 2018) Lung (Gazdhar et al, 2006;Gazdhar et al, 2007) Heart (Ayuni et al, 2010;Hargrave et al, 2014;Sugrue et al, 2020) Diaphragm (Beshay et al, 2009) Liver (Heller et al, 1996;Kobayashi et al, 2003) Tumor (Goepfert et al, 2011) Cornea (Zhou and Dean, 2007) Retina (Matsuda and Cepko, 2004;Lirong et al, 2014) Brain (Inoue and Krumlauf, 2001;De Fry et al, 2010;Nomura et al, 2016) Artery and muscle (Matsumoto et al, 2001;Zhang et al, 2001;Molnar et al, 2004;Tavakoli et al, 2006;Sokołowska and Błachnio-Zabielska, 2019) Nanochannel-based electroporation (Boukany et al, 2011;Geng and Lu, 2013;Wang and Lee, 2013;Xie et al, 2013;Gao et al, 2014;Chang et al, 2016;Gallego-Perez et al, 2016;…”

Section: Advantages Limitations Applicationsmentioning

confidence: 99%

“…More recently, Gallego-Perez et al (2017) have used the same governing physical principles to enable transfection of tissues in vivo via Tissue Nano-Transfection to induce direct cell reprogramming for regenerative applications (Gallego-Perez et al, 2017). Table 1 provides an overview of advantages and limitations for several widely used non-viral gene delivery techniques, such as electroporation (Gehl, 2003;Wells, 2004;Glover et al, 2005;Lin et al, 2005;Liu et al, 2005Liu et al, , 2006Mehier-Humbert and Guy, 2005;O'Brien and Lummis, 2006;Al-Dosari and Gao, 2009;Boukany et al, 2011;Tzeng et al, 2011;Guo and Huang, 2012;Mellott et al, 2013;Ding et al, 2014;Song et al, 2015;Das et al, 2016;Tschon et al, 2016;Tsuchiya et al, 2017;Vroomen et al, 2017;Kawai et al, 2018;Melancon et al, 2018;Shapiro et al, 2018;Shi J. et al, 2018;Pomatto et al, 2019;Tang S. et al, 2019;Bono et al, 2020), Nanochannel-based electroporation (Boukany et al, 2011;Geng and Lu, 2013;Wang and Lee, 2013;Xie et al, 2013;Gao et al, 2014;Gallego-Perez et al, 2016;Chang et al, 2016;Gallego-Perez et al, 2017;Shi J. et al, 2018), Sonoporation…”

Section: Advantages and Disadvantages Of Current Non-viral Gene Delivmentioning

confidence: 99%

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Non-viral Gene Delivery Methods for Bone and Joints

Gantenbein

Tang

Guerrero

et al. 2020

Front. Bioeng. Biotechnol.

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Viral carrier transport efficiency of gene delivery is high, depending on the type of vector. However, viral delivery poses significant safety concerns such as inefficient/unpredictable reprogramming outcomes, genomic integration, as well as unwarranted immune responses and toxicity. Thus, non-viral gene delivery methods are more feasible for translation as these allow safer delivery of genes and can modulate gene expression transiently both in vivo, ex vivo, and in vitro. Based on current studies, the efficiency of these technologies appears to be more limited, but they are appealing for clinical translation. This review presents a summary of recent advancements in orthopedics, where primarily bone and joints from the musculoskeletal apparatus were targeted. In connective tissues, which are known to have a poor healing capacity, and have a relatively low cell-density, i.e., articular cartilage, bone, and the intervertebral disk (IVD) several approaches have recently been undertaken. We provide a brief overview of the existing technologies, using nano-spheres/engineered vesicles, lipofection, and in vivo electroporation. Here, delivery for microRNA (miRNA), and silencing RNA (siRNA) and DNA plasmids will be discussed. Recent studies will be summarized that aimed to improve regeneration of these tissues, involving the delivery of bone morphogenic proteins (BMPs), such as BMP2 for improvement of bone healing. For articular cartilage/osteochondral junction, non-viral methods concentrate on targeted delivery to chondrocytes or MSCs for tissue engineering-based approaches. For the IVD, growth factors such as GDF5 or GDF6 or developmental transcription factors such as Brachyury or FOXF1 seem to be of high clinical interest. However, the most efficient method of gene transfer is still elusive, as several preclinical studies have reported many different non-viral methods and clinical translation of these techniques still needs to be validated. Here we discuss the non-viral methods applied for bone and joint and propose methods that can be promising in clinical use.

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Cell sources proposed for nucleus pulposus regeneration

et al. 2021

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Lower back pain (LBP) occurs in 80% of adults in their lifetime; resulting in LBP being one of the biggest causes of disability worldwide. Chronic LBP has been linked to the degeneration of the intervertebral disc (IVD). The current treatments for chronic back pain only provide alleviation of symptoms through pain relief, tissue removal, or spinal fusion; none of which target regenerating the degenerate IVD. As nucleus pulposus (NP) degeneration is thought to represent a key initiation site of IVD degeneration, cell therapy that specifically targets the restoration of the NP has been reviewed here. A literature search to quantitatively assess all cell types used in NP regeneration was undertaken. With key cell sources: NP cells; annulus fibrosus cells; notochordal cells; chondrocytes; bone marrow mesenchymal stromal cells; adiposederived stromal cells; and induced pluripotent stem cells extensively analyzed for their regenerative potential of the NP. This review highlights: accessibility; expansion capability in vitro; cell survival in an IVD environment; regenerative potential; and safety for these key potential cell sources. In conclusion, while several potential cell sources have been proposed, iPSC may provide the most promising regenerative potential.

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Nonviral Transfection With Brachyury Reprograms Human Intervertebral Disc Cells to a Pro‐Anabolic Anti‐Catabolic/Inflammatory Phenotype: A Proof of Concept Study

Cited by 24 publications

References 51 publications

NOTO Transcription Factor Directs Human Induced Pluripotent Stem Cell-Derived Mesendoderm Progenitors to a Notochordal Fate

NOTO Transcription Factor Directs Human Induced Pluripotent Stem Cell-Derived Mesendoderm Progenitors to a Notochordal Fate

Non-viral Gene Delivery Methods for Bone and Joints

Cell sources proposed for nucleus pulposus regeneration

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