Short regulatory DNA sequences to target brain endothelial cells for gene therapy

Müller-Fielitz, Helge; Dogbevia, Godwin; Körbelin, Jakob; Bannach, Jacqueline; Vahldieck, Carl; Kusche-Vihrog, Kristina; Müller, Oliver; Nogueiras, Rubén; Prévot, Vincent; Schwaninger, Markus

doi:10.1177/0271678x211039617

Cited by 8 publications

(5 citation statements)

References 62 publications

(89 reference statements)

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“…Further, the vector was highly endothelial-specific in this dose regime – isolated instances of neuronal or astrocytic transduction were rare ( Extended Data Figure 6 ). Notably, while both AAV-BI30 and AAV-BR1 were predominantly endothelial-directed, AAV-BI30 transduced significantly fewer non-endothelial cells than AAV-BR1 (which is known to sporadically transduce neurons and astrocytes 2 , 38 ).…”

Section: Resultsmentioning

confidence: 99%

A high-efficiency AAV for endothelial cell transduction throughout the central nervous system

et al. 2022

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Endothelial cells have a crucial role in nervous system function, and mounting evidence points to endothelial impairment as a major contributor to a wide range of neurological diseases. However, tools to genetically interrogate these cells in vivo remain limited. Here, we describe AAV-BI30, a capsid that specifically and efficiently transduces endothelial cells throughout the central nervous system. At relatively low systemic doses, this vector transduces the majority of arterial, capillary, and venous endothelial cells in the brain, retina, and spinal cord vasculature of adult C57BL/6 mice. Furthermore, we show that AAV-BI30 robustly transduces endothelial cells in multiple mouse strains and rats in vivo and human brain microvascular endothelial cells in vitro . Finally, we demonstrate AAV-BI30’s capacity to achieve efficient and endothelial-specific Cre-mediated gene manipulation in the central nervous system. This combination of attributes makes AAV-BI30 uniquely well-suited to address outstanding research questions in neurovascular biology and aid the development of therapeutics to remediate endothelial dysfunction in disease.

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Section: Resultsmentioning

confidence: 99%

A high-efficiency AAV for endothelial cell transduction throughout the central nervous system

et al. 2022

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“…Although the full-length promoter sequences in the abovementioned cases allow for gene expression in virtually all ECs, these studies prove that microenvironment-dependent transcription factors interact with respective regulatory elements on pan-endothelial promoters in an organ-specific manner. Other promoters or promoter elements seem to be induced only in distinct EC subpopulations [74]. Brain EC-specific expression of Cre recombinase driven by the Slco1c1 promoter, for example, has successfully been employed to generate a mouse model with specific gene knockout in cerebral ECs [75].…”

Section: Endothelial Heterogeneity Within the Capillary Networkmentioning

confidence: 99%

Vascular Endothelial Cells: Heterogeneity and Targeting Approaches

Hennigs

Matuszcak

Trepel³

et al. 2021

Cells

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Forming the inner layer of the vascular system, endothelial cells (ECs) facilitate a multitude of crucial physiological processes throughout the body. Vascular ECs enable the vessel wall passage of nutrients and diffusion of oxygen from the blood into adjacent cellular structures. ECs regulate vascular tone and blood coagulation as well as adhesion and transmigration of circulating cells. The multitude of EC functions is reflected by tremendous cellular diversity. Vascular ECs can form extremely tight barriers, thereby restricting the passage of xenobiotics or immune cell invasion, whereas, in other organ systems, the endothelial layer is fenestrated (e.g., glomeruli in the kidney), or discontinuous (e.g., liver sinusoids) and less dense to allow for rapid molecular exchange. ECs not only differ between organs or vascular systems, they also change along the vascular tree and specialized subpopulations of ECs can be found within the capillaries of a single organ. Molecular tools that enable selective vascular targeting are helpful to experimentally dissect the role of distinct EC populations, to improve molecular imaging and pave the way for novel treatment options for vascular diseases. This review provides an overview of endothelial diversity and highlights the most successful methods for selective targeting of distinct EC subpopulations.

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“…Here, we took a different approach, and to our knowledge this is the first The additional transduction of neurons and lung tissue observed in this study will probably only improve the treatment efficiency by increasing the NPC2 concentration in the brain and blood, respectively. To limit the transgene expression to endothelial cells of the brain, the specificity of the vector could be further improved by using an endothelial cell-specific promoter, like the regulatory sequences C-Ocln-W and C-Sls2a1-S-W, which in mice have been shown to enable a stronger and more brain endothelialspecific expression than the CAG promoter (Graßhoff et al, 2021) used in the present study. Secretion of proteins to the blood by BECs might however have limitations in other settings where the resulting protein shows off-target effects.…”

Section: Aav-br1 Gene Therapy As a Strategy For Drug Delivery To The ...mentioning

confidence: 99%

A novel strategy for deliveringNiemann‐Pick typeC2proteins across the blood–brain barrier using the brain endothelial‐specificAAV‐BR1virus

Rasmussen

Hede

Routhe

et al. 2022

Journal of Neurochemistry

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Treating central nervous system (CNS) diseases is complicated by the incapability of numerous therapeutics to cross the blood–brain barrier (BBB), mainly composed of brain endothelial cells (BECs). Genetically modifying BECs into protein factories that supply the CNS with recombinant proteins is a promising approach to overcome this hindrance, especially in genetic diseases, like Niemann Pick disease type C2 (NPC2), where both CNS and peripheral cells are affected. Here, we investigated the potential of the BEC‐specific adeno‐associated viral vector (AAV‐BR1) encoding NPC2 for expression and secretion from primary BECs cultured in an in vitro BBB model with mixed glial cells, and in healthy BALB/c mice. Transduced primary BECs had significantly increased NPC2 gene expression and secreted NPC2 after viral transduction, which significantly reversed cholesterol deposition in NPC2 deficient fibroblasts. Mice receiving an intravenous injection with AAV‐BR1‐NCP2‐eGFP were sacrificed 8 weeks later and examined for its biodistribution and transgene expression of eGFP and NPC2. AAV‐BR1‐NPC2‐eGFP was distributed mainly to the brain and lightly to the heart and lung, but did not label other organs including the liver. eGFP expression was primarily found in BECs throughout the brain but occasionally also in neurons suggesting transport of the vector across the BBB, a phenomenon also confirmed in vitro. NPC2 gene expression was up‐regulated in the brain, and recombinant NPC2 protein expression was observed in both transduced brain capillaries and neurons. Our findings show that AAV‐BR1 transduction of BECs is possible and that it may denote a promising strategy for future treatment of NPC2.

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Short regulatory DNA sequences to target brain endothelial cells for gene therapy

Cited by 8 publications

References 62 publications

A high-efficiency AAV for endothelial cell transduction throughout the central nervous system

A high-efficiency AAV for endothelial cell transduction throughout the central nervous system

Vascular Endothelial Cells: Heterogeneity and Targeting Approaches

A novel strategy for deliveringNiemann‐Pick typeC2proteins across the blood–brain barrier using the brain endothelial‐specificAAV‐BR1virus

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