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Shattuck-Brandt, R.

Publications and source records attributed to Shattuck-Brandt, R..

2 recordsLinked to original sources

Amyloid-β-regulated gene circuits for programmable Alzheimer's disease therapy

Alzheimer's disease (AD) is a neurodegenerative disease characterized in part by the accumulation of the protein amyloid-{beta} (A{beta}). Monoclonal antibodies (mAbs) that target A{beta} for clearance from the brain have received FDA approval; however, these therapies are accompanied by serious side effects, and their cognitive benefit for patients remains of tremendous debate. Here, we present a potential engineered cell therapy for AD in which we enlist cells of the central nervous system as programmable agents for sculpting the neurodegenerative niche toward one that mitigates glial reactivity and neuronal loss. We constructed a suite of A{beta}-sensitive synthetic Notch (synNotch) receptors from clinically tested anti-A{beta} mAbs and show that cells expressing these receptors can recognize synthetic A{beta}42 and A{beta}40 with differential sensitivity. We express these receptors in astrocytes, cells native to the brain that are known to become dysfunctional in AD. These synNotch astrocytes, which upregulate selected transgenes upon exposure to synthetic and human brain-derived amyloid, were engineered to express potential therapeutic transgenes in response to A{beta}, including brain-derived neurotrophic factor and antagonists of the cytokines tumor necrosis factor and interleukin-1. SynNotch astrocytes that express such antagonists in response to A{beta} partially attenuate a cytokine-induced reactive astrocyte phenotype and promote barrier properties in brain microvascular endothelial cells. Additionally, engineered A{beta}-synNotch cells potently upregulate transgene expression in response to A{beta} deposited in the 5xFAD mouse brain, demonstrating the capacity to recognize A{beta} in situ. Overall, our work supports A{beta}-synNotch receptors as promising tools to generate a cell-based therapy for AD with targeted functionalities to positively influence the AD niche.

bioengineering↗

A programmable arthritis-specific receptor for guided articular cartilage regenerative medicine

ObjectiveInvestigational cell therapies have been developed as disease-modifying agents for the treatment of osteoarthritis (OA), including those that inducibly respond to inflammatory factors driving OA progression. However, dysregulated inflammatory cascades do not specifically signify the presence of OA. Here, we deploy a synthetic receptor platform that regulates cell behaviors in an arthritis-specific fashion to confine transgene expression to sites characterized by cartilage degeneration. MethodsAn scFv specific for type II collagen (CII) was used to produce a synthetic Notch (synNotch) receptor that enables "CII-synNotch" mesenchymal stromal cells (MSCs) to recognize CII fibers exposed in damaged cartilage. Engineered cell activation by both CII-treated culture surfaces and on primary tissue samples was measured via inducible reporter transgene expression. TGF{beta}3-expressing cells were assessed for cartilage anabolic gene expression via qRT-PCR. In a co-culture with CII-synNotch MSCs engineered to express IL-1Ra, ATDC5 chondrocytes were stimulated with IL-1, and inflammatory responses of ATDC5s were profiled via qRT-PCR and an NF-kB reporter assay. ResultsCII-synNotch MSCs are highly responsive to CII, displaying activation ranges over 40-fold in response to physiologic CII inputs. CII-synNotch cells exhibit the capacity to distinguish between healthy and damaged cartilage tissue and constrain transgene expression to regions of exposed CII fibers. Receptor-regulated TGF{beta}3 expression resulted in upregulation of Acan and Col2a1 in MSCs, and inducible IL-1Ra expression by engineered CII-synNotch MSCs reduced pro-inflammatory gene expression in chondrocytes. ConclusionThis work demonstrates proof-of-concept that the synNotch platform guides MSCs for spatially regulated, disease-dependent delivery of OA-relevant biologic drugs.

bioengineering↗