Lentiviral vectors carrying enhancer elements of Hb9 promoter drive selective transgene expression in mouse spinal cord motor neurons

Peviani, Marco; Kurosaki, Mami; Terao, Mineko; Lidonnici, D.; Gensano, Francesco; Battaglia, Elisa; Tortarolo, Massimo; Piva, Roberto; Bendotti, Caterina

doi:10.1016/j.jneumeth.2011.12.024

Cited by 21 publications

(23 citation statements)

References 39 publications

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“…The proximal sequences from nt −500 to 0 are necessary for binding Nkx2.2 and Irx3, which repress HB9 in interneurons (Lee et al, 2004). Lack of the proximal HB9 promoter sequences may thus contribute to LvHB9::RFP expression in human iPSc-derived interneurons, in line with data from chicken and mouse (Lee et al, 2004;Nakano et al, 2005;Peviani et al, 2012).…”

Section: Discussionsupporting

confidence: 62%

Modeling amyotrophic lateral sclerosis in pure human iPSc-derived motor neurons isolated by a novel FACS double selection technique

Toli

Buttigieg

Blanchard

et al. 2015

Neurobiology of Disease

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Section: Discussionsupporting

confidence: 62%

Modeling amyotrophic lateral sclerosis in pure human iPSc-derived motor neurons isolated by a novel FACS double selection technique

Toli

Buttigieg

Blanchard

et al. 2015

Neurobiology of Disease

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“…Given that AAV vectors has limited cloning capacity (i.e., large plasmids can hardly be packaged into AAV viral particles), we have chosen a genetically engineered minimal promoter of the Hb9 gene (Hb9_AB), which is about 550 bp in length and has already been demonstrated to specifically drive the expression of a transgene into MNs, both in vitro and in vivo 29 . For the preparation of the viral particles, we have chosen the AAV9 serotype because of its broad tropism, including the capacity to target neurons 28 .…”

Section: Discussionmentioning

confidence: 99%

“…cytosol, mitochondrial matrix and ER lumen 13, 14, 19, 20 under the control of either a minimal promoter (Hb9_AB) derived from the (MN)-specific promoter of the Hb9 gene 29 , or the pan-neuronal hSyn promoter.…”

Section: Methodsmentioning

confidence: 99%

“…Subsequently, the [hSyn] promoter in the obtained plasmids was substituted with the sequence coding for the ∼550 bp MN specific promoter [Hb9_AB], which was previously amplified by PCR using the Bg.Hb9_AB_GFP plasmid 29 (kindly provided by Dr. Caterina Bendotti, IRCCS Istituto di Ricerche Farmacologiche Mario Negri, Milano, Italy) as template, and primers AB-MluI-F and AB-KpnI-R (sequences are reported in the Supplementary Information). The PCR product, subjected to MluI-KpnI cleavage and purified, was then ligated into both pAAV-[hSyn]-D1cpv and pAAV-[hSyn]-D4ERcpv plasmidic fragments digested with the same enzymes.…”

Section: Methodsmentioning

confidence: 99%

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Generation and validation of novel adeno-associated viral vectors for the analysis of Ca2+ homeostasis in motor neurons

Norante

Massimino

Lorenzon

et al. 2017

Sci Rep

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A finely tuned Ca2+ homeostasis in restricted cell domains is of fundamental importance for neurons, where transient Ca2+ oscillations direct the proper coordination of electro-chemical signals and overall neuronal metabolism. Once such a precise regulation is unbalanced, however, neuronal functions and viability are severely compromised. Accordingly, disturbed Ca2+ metabolism has often been claimed as a major contributor to different neurodegenerative disorders, such as amyotrophic lateral sclerosis that is characterised by selective motor neuron (MN) damage. This notion highlights the need for probes for the specific and precise analysis of local Ca2+ dynamics in MNs. Here, we generated and functionally validated adeno-associated viral vectors for the expression of gene-encoded fluorescent Ca2+ indicators targeted to different cell domains, under the transcriptional control of a MN-specific promoter. We demonstrated that the probes are specifically expressed, and allow reliable local Ca2+ measurements, in MNs from murine primary spinal cord cultures, and can also be expressed in spinal cord MNs in vivo, upon systemic administration to newborn mice. Preliminary analyses using these novel vectors have shown larger cytosolic Ca2+ responses following stimulation of AMPA receptors in the cytosol of primary cultured MNs from a murine genetic model of ALS compared to the healthy counterpart.

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“…Later studies reported that inclusion of NSE promoter achieved relatively precise but not exclusive neuronal expression (Navarro et al, 1999; Kügler et al, 2003). Among uncommon promoters, LV encoding a 1.6 kb segment of ‘homeobox 9’ promoter was used for motor neuron targeting in mice spinal cord (Peviani et al, 2012). Also, Thy1.2 and α-CAMKII promoter were shown to be specific for pyramidal neurons (Dittgen et al, 2004).…”

Section: Cns-targeted Gene Delivery Using Cell-specific Promotersmentioning

confidence: 99%

Destination Brain: the Past, Present, and Future of Therapeutic Gene Delivery

Joshi

Labhasetwar

Ghorpade

2017

J Neuroimmune Pharmacol

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Neurological diseases and disorders (NDDs) present a significant societal burden and currently available drug- and biological-based therapeutic strategies have proven inadequate to alleviate it. Gene therapy is a suitable alternative to treat NDDs compared to conventional systems since it can be tailored to specifically alter select gene expression, reverse disease phenotype and restore normal function. The scope of gene therapy has broadened over the years with the advent of RNA interference and genome editing technologies. Consequently, encouraging results from central nervous system (CNS)-targeted gene delivery studies have led to their transition from preclinical to clinical trials. As we shift to an exciting gene therapy era, a retrospective of available literature on CNS-associated gene delivery is in order. This review is timely in this regard, since it analyzes key challenges and major findings from the last two decades and evaluates future prospects of brain gene delivery. We emphasize major areas consisting of physiological and pharmacological challenges in gene therapy, function-based selection of an ideal cellular target, available therapy modalities, and diversity of viral vectors and nanoparticles as vehicle systems. Further, we present plausible answers to key questions such as strategies to circumvent low blood-brain barrier permeability, most suitable CNS cell types for targeting. We compare and contrast pros and cons of the tested viral vectors in context of delivery systems used in past and current clinical trials. Gene vector design challenges are also evaluated in the context of cell-specific promoters. Key challenges and findings reported for recent gene therapy clinical trials, assessing viral vectors and nanoparticles, are discussed in the context of bench to bedside gene therapy translation. We conclude this review by tying together gene delivery challenges, available vehicle systems and comprehensive analysis of neuropathogenesis to outline future prospects of CNS-targeted gene therapies.

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Lentiviral vectors carrying enhancer elements of Hb9 promoter drive selective transgene expression in mouse spinal cord motor neurons

Cited by 21 publications

References 39 publications

Modeling amyotrophic lateral sclerosis in pure human iPSc-derived motor neurons isolated by a novel FACS double selection technique

Modeling amyotrophic lateral sclerosis in pure human iPSc-derived motor neurons isolated by a novel FACS double selection technique

Generation and validation of novel adeno-associated viral vectors for the analysis of Ca2+ homeostasis in motor neurons

Destination Brain: the Past, Present, and Future of Therapeutic Gene Delivery

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