A5080340349- CAR-T cell therapy research
- CRISPR and Genetic Engineering
- Virus-based gene therapy research
- Immune Cell Function and Interaction
- Nanowire Synthesis and Applications
- RNA Interference and Gene Delivery
- Ubiquitin and proteasome pathways
- Advanced biosensing and bioanalysis techniques
- Retinal Imaging and Analysis
- Retinal Diseases and Treatments
- vaccines and immunoinformatics approaches
- Protein Degradation and Inhibitors
- Axon Guidance and Neuronal Signaling
- Immune Response and Inflammation
- Endoplasmic Reticulum Stress and Disease
- Viral Infectious Diseases and Gene Expression in Insects
- Glaucoma and retinal disorders
- Signaling Pathways in Disease
- Multiple Myeloma Research and Treatments
- Microfluidic and Bio-sensing Technologies
- SARS-CoV-2 and COVID-19 Research
- Hippo pathway signaling and YAP/TAZ
- Cancer-related molecular mechanisms research
- Cancer-related gene regulation
- Retinal and Optic Conditions
University of California, San Francisco
2022-2025
Gladstone Institutes
2022-2025
Karolinska University Hospital
2016-2025
Karolinska Institutet
2016-2025
Parker Institute for Cancer Immunotherapy
2022
Queensland University of Technology
2015
Penn Presbyterian Medical Center
1993
The efficacy of adoptive T cell therapies for cancer treatment can be limited by suppressive signals from both extrinsic factors and intrinsic inhibitory checkpoints1,2. Targeted gene editing has the potential to overcome these limitations enhance therapeutic function3-10. Here we performed multiple genome-wide CRISPR knock-out screens under different immunosuppressive conditions identify genes that targeted prevent dysfunction. These converged on RASA2, a RAS GTPase-activating protein...
Precise targeting of large transgenes to T cells using homology-directed repair has been transformative for adoptive cell therapies and biology. Delivery DNA templates via adeno-associated virus (AAV) greatly improved knockin efficiencies, but the tropism current AAV serotypes restricts their use human employed in immunodeficient mouse models. To enable targeted knockins murine cells, we evolved Ark313, a synthetic that exhibits high transduction efficiency cells. We performed genome-wide...
Chronic stimulation can cause T cell dysfunction and limit the efficacy of cellular immunotherapies. Improved methods are required to compare large numbers synthetic knockin (KI) sequences reprogram functions. Here, we developed modular pooled KI screening (ModPoKI), an adaptable platform for construction DNA libraries using barcoded multicistronic adaptors. We built two ModPoKI 100 transcription factors (TFs) 129 natural surface receptors (SRs). Over 30 screens across human TCR- CAR-T cells...
Abstract Engineering T cell specificity and function at multiple loci can generate more effective cellular therapies, but current manufacturing methods produce heterogenous mixtures of partially engineered cells. Here we develop a one-step process to enrich unlabeled cells containing knock-ins target using family repair templates named synthetic exon expression disruptors (SEEDs). SEEDs associate transgene integration with the disruption paired endogenous surface protein while preserving in...
There is currently a lack of tools capable perturbing genes in both precise and spatiotemporal fashion. The flexibility CRISPR (clustered regularly interspaced short palindromic repeats), coupled with light's unparalleled resolution deliverable from controllable source, makes optogenetic well-suited solution for gene perturbations. Here, we present new tool (Blue Light-inducible Universal VPR-Improved Production RGRs, BLU-VIPR) that diverges prevailing split-Cas design strategies instead...
Endoplasmic reticulum (ER) stress is a physiological response to protein overload or misfolded proteins in the ER. Certain anti-cancer drugs, e.g. bortezomib and nelfinavir, induce ER implying that this could be successful therapeutic strategy against several forms of cancer. To find novel ER-stress inducers we screened panel natural synthetic Toll-like receptor (TLR) agonists human keratinocytes identified drug imiquimod (IMQ) as potent inducer stress. Other TLR7 TLR8 agonists, including...
TRIM21 is an interferon-stimulated E3 ligase that controls the activity of pattern-recognition signaling via ubiquitination interferon regulatory factors and DDX41. Previous studies on role in innate immune responses have yielded contradictory results, suggesting cell specific. Here, we report bone-marrow-derived macrophages (BMDMs) generated from Trim21-/- mice reduced expression mature macrophage markers. Reflecting their differentiation response to colony-stimulating factor (M-CSF), BMDMs...
Neonatal lupus erythematosus (NLE) may develop after transplacental transfer of maternal autoantibodies with cardiac manifestations (congenital heart block, CHB) including atrioventricular atrial and ventricular arrhythmias, cardiomyopathies. The association anti-Ro/SSA antibodies is well established, but a recurrence rate only 12%-16% despite persisting suggests that additional factors are required for CHB development. Here, we identify fetal genetic variants conferring risk elucidate their...
Abstract Systemic autoimmune diseases are characterized by the overexpression of type I IFN stimulated genes, and accumulating evidence indicate a role for IFNs in these diseases. However, underlying mechanisms this still poorly understood. To explore regulated miRNAs systemic disease, we cellular expression during both acute chronic responses. We identified T cell‐specific reduction miR‐31‐5p levels, after intramuscular injection IFNβ patients with Sjögren's syndrome (SjS). interrogate...
Abstract Multiplexed reprogramming of T cell specificity and function can generate powerful next-generation cellular therapies. However, current manufacturing methods produce heterogenous mixtures partially engineered cells. Here, we develop a one-step process to enrich for unlabeled cells with knock-ins at multiple target loci using family repair templates named S ynthetic E xon/ xpression Disruptors (SEEDs). SEED engineering associates transgene integration the disruption paired endogenous...
SUMMARY Chronic stimulation can cause T cell dysfunction and limit efficacy of cellular immunotherapies. CRISPR screens have nominated gene targets for engineered cells, but improved methods are required to compare large numbers synthetic knockin sequences reprogram functions. Here, we developed Modular Pooled Knockin Screening (ModPoKI), an adaptable platform modular construction DNA libraries using barcoded multicistronic adaptors. We built two ModPoKI 100 transcription factors (TFs) 129...
Abstract Melanoma tumors are highly metastatic partly due to the ability of melanoma cells transition between invasive and proliferative states. However, mechanisms underlying this plasticity still not fully understood. To identify new epigenetic regulators plasticity, we combined data mining, tumor models, proximity proteomics, CUT&RUN sequencing. We focus on druggable family bromodomain readers TRIM28 as a regulator plasticity. find that promotes expression pro‐invasive genes controls...
ABSTRACT Adoptive chimeric antigen receptor T-cell (CAR-T) therapy is transformative and approved for hematologic malignancies. It also being developed the treatment of solid tumors, autoimmune disorders, heart disease, aging. Despite unprecedented clinical outcomes, CAR-T other engineered cell therapies face a variety manufacturing safety challenges. Traditional methods, such as lentivirus transduction electroporation, result in random integration or cause significant cellular damage, which...
Discovering the role of fibroblasts residing in tumor microenvironment (TME) requires controlled, localized perturbations because play critical roles regulating immunity and biology at multiple sites. Systemic can lead to unintended, confounding secondary effects, methods locally genetically engineer are lacking. To specifically investigate murine stromal cell restricted TME, we developed an adeno-associated virus (AAV)-based method target any gene-of-interest high efficiency (>80%). As...
<title>Abstract</title> Adoptive chimeric antigen receptor T-cell (CAR-T) therapy is transformative and approved for hematologic malignancies. It also being developed the treatment of solid tumors, autoimmune disorders, heart disease, aging. Despite unprecedented clinical outcomes, CAR-T other engineered cell therapies face a variety manufacturing safety challenges. Traditional methods, such as lentivirus transduction electroporation, result in random integration or cause significant...
To localize choroidal lesions demonstrated by indocyanine green (ICG) angiography, retinal blood vessels must be identified. Since are seen in early-phase ICG angiography and the best on late-phase images, a technique has been developed to demonstrate late phase of study. The requires injection 0.3 mg dye mixed 3.0 cc diluent. This is given between 30 40 minutes into angiogram. allows visualization vessels, which can used as landmarks lesions. Additionally, this technique, when combined with...
The introduction of immune checkpoint blockade has revolutionized the treatment metastatic melanoma 1 . However, 40-60% patients with do not respond to blockade, and a significant fraction acquire resistance 2,3 This resilience aggressiveness tumors is partly due their ability switch between invasive proliferative states 4,5 transition phenotypic indicates that phenotype switching occurs through reversible epigenetic mechanisms rather than by acquisition mutations 6,7 Identifying underlie...
There is currently a lack of tools capable perturbing genes in both precise and spatiotemporal fashion. CRISPR's ease use flexibility, coupled with light's unparalleled resolution deliverable from controllable source, makes optogenetic CRISPR well-suited solution for gene perturbations. Here we present new tool, BLU-VIPR, that diverges prevailing split-Cas design strategies instead focuses on regulation gRNA production. This simplifies perturbation works vivo cells previously intractable to...