Cellular fusion for gene delivery to SCA1 affected Purkinje neurons

Chen, K. Amy; Cruz, Pedro; Lanuto, Derek J.; Flotte, Terence R.; Borchelt, David R.; Srivastava, Arun; Zhang, Jianyi; Steindler, Dennis A.; Zheng, Tong

doi:10.1016/j.mcn.2011.03.003

Cited by 31 publications

(29 citation statements)

References 73 publications

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“…Moreover, we observed fusion occurring within 2 weeks after the hfMSC transplantation into the cerebella, which is far earlier than the fusion resulting from intravenous injection of bone marrow derived cells (BMDCs) (at least 24 weeks) [48]. …”

Section: Discussionmentioning

confidence: 99%

“…The fusion of peripheral stem cells with PCs has mainly been studied using BMDCs [22,48,50–53]. For example, Alvarez-Dolado et al have convincingly demonstrated fusion of BMDCs with PCs through transplantation of BMDCs expressing Cre and GFP into the peripheral circulation of lethally irradiated R26R transgenic mice expressing LacZ after Cre/lox recombination, although the frequency was very low [48,50].…”

Section: Discussionmentioning

confidence: 99%

“…For example, Alvarez-Dolado et al have convincingly demonstrated fusion of BMDCs with PCs through transplantation of BMDCs expressing Cre and GFP into the peripheral circulation of lethally irradiated R26R transgenic mice expressing LacZ after Cre/lox recombination, although the frequency was very low [48,50]. Later, Magrassi et al [22] showed that the number of PCs fusing with BMDCs in mice increases by 10 times when PCs are pharmacologically degenerated, thus suggesting the increased fusogenic potential of degenerating PCs.…”

Section: Discussionmentioning

confidence: 99%

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Fusion of Human Fetal Mesenchymal Stem Cells with “Degenerating” Cerebellar Neurons in Spinocerebellar Ataxia Type 1 Model Mice

et al. 2016

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Mesenchymal stem cells (MSCs) migrate to damaged tissues, where they participate in tissue repair. Human fetal MSCs (hfMSCs), compared with adult MSCs, have higher proliferation rates, a greater differentiation capacity and longer telomeres with reduced senescence. Therefore, transplantation of quality controlled hfMSCs is a promising therapeutic intervention. Previous studies have shown that intravenous or intracortical injections of MSCs result in the emergence of binucleated cerebellar Purkinje cells (PCs) containing an MSC-derived marker protein in mice, thus suggesting a fusion event. However, transdifferentiation of MSCs into PCs or transfer of a marker protein from an MSC to a PC cannot be ruled out. In this study, we unequivocally demonstrated the fusion of hfMSCs with murine PCs through a tetracycline-regulated (Tet-off) system with or without a Cre-dependent genetic inversion switch (flip-excision; FLEx). In the FLEx-Tet system, we performed intra-cerebellar injection of viral vectors expressing tetracycline transactivator (tTA) and Cre recombinase into either non-symptomatic (4-week-old) or clearly symptomatic (6–8-month-old) spinocerebellar ataxia type 1 (SCA1) mice. Then, the mice received an injection of 50,000 genetically engineered hfMSCs that expressed GFP only in the presence of Cre recombinase and tTA. We observed a significant emergence of GFP-expressing PCs and interneurons in symptomatic, but not non-symptomatic, SCA1 mice 2 weeks after the MSC injection. These results, together with the results obtained using age-matched wild-type mice, led us to conclude that hfMSCs have the potential to preferentially fuse with degenerating PCs and interneurons but not with healthy neurons.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Fusion of Human Fetal Mesenchymal Stem Cells with “Degenerating” Cerebellar Neurons in Spinocerebellar Ataxia Type 1 Model Mice

et al. 2016

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“…This is within the same tumor as well, creating opportunities for CSC populations with different CD markers to occur within the same tumor, making isolation and treatment of CSCs very arduous if based on these markers alone. The fusion of bone marrow derived cells (BMDCs) have been shown to occur with Purkinje neurons, cardiomyomycytes, and hepatocytes with evidence that trans-differentiation occurred only in these fused cells (AlvarezDolado et al, 2003;Chen et al, 2011). Trans-differentiation is a key characteristic of many tumors Shekhani et al, 2013).…”

Section: Theories Of Carcinogenesismentioning

confidence: 99%

Microbial influences on hormesis, oncogenesis, and therapy: A review of the literature

Clanton

Saucier

Ford

et al. 2015

Environmental Research

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“…Thus far, approaches have been based on surgical reconnection of transected mammalian nerve fibres, combined with the application of polymers such as polyethylene glycol (PEG) [72][73][74][75][76][77] . The mechanism through which PEG acts is not clear, but it is proposed to work like a fusogen by facilitating membrane merging 72 .…”

Section: Discussionmentioning

confidence: 99%

Function and regulation of the fusogen EFF-1 during axonal repair

Linton¹

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In the current clinical environment, treatment options for nerve and spinal cord injuries remain limited. To develop novel therapies, we require a better understanding of axonal repair and its underlying molecular mechanisms. The response of an axon to a transection injury is complex: the axon still attached to the cell body begins a process of regenerative regrowth, whereas a concurrent process of degeneration is activated in the severed axonal fragment. Characterizing ways in which the axon can achieve repair, involving both regeneration and degeneration, is crucial to making progress in a clinical setting.We have studied a means of axonal repair called axonal fusion, which occurs spontaneously in the mechanosensory neurons of the nematode C. elegans. Following axotomy with a UV laser, the regrowing axonal fragment is able to directly fuse with its own severed fragment, re-establishing continuity. When axonal fusion was first described in C. elegans, the molecular pathways involved were unknown, and the only protein associated with the process was the fusogen Epithelial Fusion Failure-1 (EFF-1). EFF-1 is a nematode-specific transmembrane glycoprotein that acts to fuse plasma membranes. It has been studied extensively in other C. elegans tissues, but its function and regulation in neurons have been largely uncharacterized.In this context, we aimed to further understand the role of EFF-1 in axonal repair, focusing on its potential contribution to both regeneration and degeneration, and the mechanisms that regulate its fusogenic activity. We approached these biological questions using a combination of genetic and molecular biology techniques available in the C. elegans model system.The following chapters characterize EFF-1 function and regulation in the C. elegans mechanosensory neuron PLM. Firstly, we demonstrate a cell-autonomous role for EFF-1 in axonal fusion, and show that it has a dynamic localization pattern in the regenerating axon, whereby it is mobilized to the membrane of the regenerating growth cone. We also place it downstream in a pathway of apoptotic clearance molecules that allow recognition of the distal axonal fragment.Secondly, we find that neuronal EFF-1 is regulated by the endocytic GTPase RAB-5, with alterations in RAB-5 activity affecting both EFF-1 localization and its function in axonal fusion.Thirdly, we characterize the genes involved in the degeneration of the distal PLM axon following axotomy, and find a remarkable overlap with the molecules involved in regenerative fusion, possibly including EFF-1. Finally, we discuss an intriguing potential role for EFF-1 in mediating neuronal repair through cell-cell fusion. The research detailed here represents significant progress in understanding how a fusogen mediates axonal repair in vivo. It will potentially contribute to the application of axonal fusion as a novel therapy for patients with nerve injuries.ii Declaration by authorThis thesis is composed of my original work, and contains no material previously published or written by an...

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Cellular fusion for gene delivery to SCA1 affected Purkinje neurons

Cited by 31 publications

References 73 publications

Fusion of Human Fetal Mesenchymal Stem Cells with “Degenerating” Cerebellar Neurons in Spinocerebellar Ataxia Type 1 Model Mice

Fusion of Human Fetal Mesenchymal Stem Cells with “Degenerating” Cerebellar Neurons in Spinocerebellar Ataxia Type 1 Model Mice

Microbial influences on hormesis, oncogenesis, and therapy: A review of the literature

Function and regulation of the fusogen EFF-1 during axonal repair

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