Radin, D. | bioRxiv Science

bioRxiv2023

Intranasal Delivery of shRNA to Knockdown the 5HT-2A Receptor Enhances Memory and Alleviates Anxiety

Short-hairpin RNAs (shRNA) targeting knockdown of specific genes hold enormous promise for precision-based therapeutics to treat numerous neurodegenerative disorders. However, whether shRNA constructed molecules can modify neuronal circuits underlying certain behaviors has not been explored. We designed shRNA to knockdown the human HTR2A gene in vitro using iPSC-differentiated neurons. Multi-electrode array (MEA) results showed the knockdown of the 5HT-2A mRNA and receptor protein led to a decrease in spontaneous electrical activity. In vivo, intranasal delivery of AAV9 vectors containing shRNA resulted in a decrease in anxiety-like behavior in mice and a significant improvement in memory in both mice (104%) and rats (92%) compared to vehicle-treated animals. Our demonstration of a non-invasive shRNA delivery platform that can bypass the blood-brain barrier has broad implications for treating numerous neurological mental disorders. Specifically, targeting the HTR2A gene presents a novel therapeutic approach for treating chronic anxiety and age-related cognitive decline.

neuroscience↗

bioRxiv2020

A Microglial Subset at the Tumor-Stroma Interface of Glioma

BackgroundMyeloid involvement in High Grade Gliomas, such as Glioblastoma, has become apparent and detrimental to disease outcomes. There is great interest in characterizing the HGG tumor microenvironment to understand how neoplastic lesions are supported, and to devise novel therapeutic targets. The tumor microenvironment of the central nervous system is unique as it contains neural and specialized glial cells, including the resident myeloid cells, microglia. Glioma-associated microglia and peripherally infiltrating monocytes/macrophages (GAM) accumulate within the neoplastic lesion where they facilitate tumor growth and drive immunosuppression. A longstanding limitation has been the ability to accurately differentiate microglia and macrophage roles in pathology, and identify the consequences of the spatial organization of these cells. ResultsHere we characterize the tumor-stroma border and identify peripheral glioma-associated microglia (PGAM) at the tumor leading edge as a unique subpopulation of GAM. Using data mining and analyses of samples derived from both murine and human sources, we show that PGAM exhibit a pro-inflammatory and chemotactic phenotype that is associated with peripheral monocyte recruitment, poorly enhancing radiomic features, and decreased overall survival. ConclusionsPGAM act as a unique subset of GAM, at the tumor-stroma interface, corresponding to disease outcomes. We propose the application of a novel gene signature to identify these cells, and suggest that PGAM constitute a cellular target of the TME. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=126 SRC="FIGDIR/small/357459v1_ufig1.gif" ALT="Figure 1"> View larger version (76K): org.highwire.dtl.DTLVardef@6c08b6org.highwire.dtl.DTLVardef@b01421org.highwire.dtl.DTLVardef@1f99f9aorg.highwire.dtl.DTLVardef@15d5011_HPS_FORMAT_FIGEXP M_FIG C_FIG

neuroscience↗