A5051765438- CAR-T cell therapy research
- Chemical Synthesis and Analysis
- RNA and protein synthesis mechanisms
- Microbial Natural Products and Biosynthesis
- Nanowire Synthesis and Applications
- Monoclonal and Polyclonal Antibodies Research
- Viral Infectious Diseases and Gene Expression in Insects
- Biochemical and Structural Characterization
- Click Chemistry and Applications
- Virus-based gene therapy research
- Biosimilars and Bioanalytical Methods
- Ubiquitin and proteasome pathways
- Protein Degradation and Inhibitors
- Immune Cell Function and Interaction
- Phosphodiesterase function and regulation
- Enzyme Catalysis and Immobilization
- T-cell and B-cell Immunology
- Radiopharmaceutical Chemistry and Applications
- Escherichia coli research studies
- Immune Response and Inflammation
- Acoustic Wave Resonator Technologies
- Prostate Cancer Treatment and Research
- Synthesis and Catalytic Reactions
- Nanofabrication and Lithography Techniques
- Antimicrobial Peptides and Activities
California Institute for Biomedical Research
2015-2023
Scripps Research Institute
2009-2023
Scripps (United States)
2020-2022
Scripps Institution of Oceanography
2020-2022
Torrey Pines Institute For Molecular Studies
2015
Harvard University
2011-2013
Genomics Institute of the Novartis Research Foundation
2009
University of Wisconsin–Madison
2002
Targeting STING for cancer therapy Activation of the (stimulator interferon genes) protein by cyclic dinucleotide metabolites plays a critical role in antitumor immunity. The development synthetic agonists is therefore being pursued as strategy therapy, but inherent instability dinucleotides has limited current efforts. Pan et al. and Chin identified stable that act “closed” conformation similar to natural ligand, guanosine monophosphate–adenosine monophosphate (see Perspective Gajewski...
Significance Chimeric antigen receptor T (CAR-T) cell therapy has produced promising results in clinical trials but been challenged by the inability to control engineered cells once infused into patient. Here we present a generalizable method of controlling CAR-T using peptide-engrafted antibody-based molecular switches that act as bridge between target and cell. We show specific for CD19 govern activity, tissue-homing, cytokine release, phenotype switchable dose-titratable manner xenograft...
The adoptive transfer of autologous T cells engineered to express a chimeric antigen receptor (CAR) has emerged as promising cancer therapy. Despite impressive clinical efficacy, the general application current CAR-T--cell therapy is limited by serious treatment-related toxicities. One approach improve safety CAR-T involves making their activation and proliferation dependent upon adaptor molecules that mediate formation immunological synapse between target cell T-cell. Here, we describe...
We have employed a rapid fluorescence-based screen to assess the polyspecificity of several aminoacyl-tRNA synthetases (aaRSs) against an array unnatural amino acids. discovered that p-cyanophenylalanine specific synthetase (pCNF-RS) has high substrate permissivity for acids, while maintaining its ability discriminate 20 canonical This orthogonal pCNF-RS, together with cognate amber nonsense suppressor tRNA, is able selectively incorporate 18 acids into proteins, including trifluoroketone-,...
Significance This work describes a facile system for incorporating noncanonical amino acids containing long side-chain thiols using an expanded genetic code. These begin to overcome the distance and geometric constraints of cysteine disulfide can pair with cysteines cross-link more remote sites in proteins. To demonstrate this notion, we constructed library random β-lactamase mutants these grew them at nonpermissive temperatures. We identified mutant enzyme that is cross-linked by one such...
Chimeric antigen receptor (CAR)-engineered T cells (CAR-Ts) provide a potent antitumor response and have become promising treatment option for cancer. However, despite their efficacy, CAR-T are associated with significant safety challenges related to the inability control activation expansion terminate response. Herein, we demonstrate that bifunctional small molecule "switch" consisting of folate conjugated fluorescein isothiocyanate (folate-FITC) can redirect regulate FITC-specific cell...
Objective Pancreatic ductal adenocarcinoma (PDAC) is a disease of unmet medical need. While immunotherapy with chimeric antigen receptor T (CAR-T) cells has shown much promise in haematological malignancies, their efficacy for solid tumours challenged by the lack tumour-specific antigens required to avoid on-target, off-tumour effects. Switchable CAR-T whereby activity cell controlled dosage tumour antigen-specific recombinant Fab-based ‘switch’ afford fully tunable response may overcome...
We report a bacterial system for the evolution of cyclic peptides that makes use an expanded set amino acid building blocks. Orthogonal aminoacyl-tRNA synthetase/tRNA CUA pairs, together with split intein were used to biosynthesize library ribosomal containing acids unique structures and reactivities. This peptide was subsequently evolve inhibitor HIV protease using selection based on cellular viability. Two three isolated after two rounds contained keto p -benzoylphenylalanine ( BzF). The...
The cyanobactin ribosomal peptide (RP) natural product pathway was manipulated to incorporate multiple tandem mutations and non-proteinogenic amino acids, using eight heterologous components simultaneously expressed in Escherichia coli . These studies reveal the potential of RPs for rational synthesis complex, new small molecules over multiple-step biosynthetic pathways simple genetic engineering.
Abstract Chimeric antigen receptor T (CAR‐T) cells have demonstrated promising results against hematological malignancies, but encountered significant challenges in translation to solid tumors. To overcome these hurdles, we developed a switchable CAR‐T cell platform which the activity of engineered is controlled by dosage an antibody‐based switch. Herein, apply this approach Her2‐expressing breast cancers engineering switch molecules through site‐specific incorporation FITC or grafting...
We have developed a novel antibody-drug conjugate (ADC) that can selectively deliver the Lck inhibitor dasatinib to human T lymphocytes. This ADC is based on humanized antibody binds with high affinity CXCR4, an antigen expressed hematopoietic cells. The resulting dasatinib-antibody suppresses T-cell-receptor (TCR)-mediated T-cell activation and cytokine expression low nM EC50 has minimal effects cell viability. may lead new class of selective immunosuppressive drugs improved safety extend...
New drugs are needed to treat gram-negative bacterial infections. These bacteria protected by an outer membrane which prevents many antibiotics from reaching their cellular targets. The leaflet of the contains LPS, is responsible for creating this permeability barrier. Interfering with LPS biogenesis affects viability. We developed a cell-based screen that identifies inhibitors biosynthesis and transport exploiting nonessentiality pathway in Acinetobacter used find inhibitor MsbA,...
Significance Chimeric antigen receptor (CAR) T cell therapy represents a powerful strategy in immuno-oncology. Nevertheless, associated life-threatening toxicities and chronic B aplasia have underscored the need to control engineered cells patient. To address these challenges, we previously developed switchable CAR (sCAR) platform that allows dose-titratable over activity by using antibody-based switches. Here, demonstrate syngeneic murine model can impart antitumor efficacy while...
Transcriptional coregulators, which mediate chromatin-dependent transcriptional signaling, represent tractable targets to modulate tumorigenic gene expression programs with small molecules. Genetic loss-of-function studies have recently implicated the coactivator, ENL, as a selective requirement for survival of acute leukemia and highlighted an essential role its chromatin reader YEATS domain. Motivated by these discoveries, we executed screen nearly 300,000 molecules identified...
The treatment of patients with acute myeloid leukemia (AML) targeted immunotherapy is challenged by the heterogeneity disease and a lack tumor-exclusive antigens. Conventional targets for AML such as CD33 CD123 have been proposed chimeric antigen receptor (CAR)-engineered T-cells (CAR-T-cells), therapy that has highly successful in B-cell lymphoma. However, are present on hematopoietic stem cells, targeting CAR-T-cells potential to elicit long-term myelosuppression. C-type lectin-like...
Significance Thiopeptides are a subclass of ribosomally synthesized natural products with complex structures and potent antimicrobial activities. Here we describe general strategy that allows the incorporation noncanonical amino acids into thiopeptides by introducing orthogonal amber suppressor aminoacyl-tRNA synthetase/tRNA pairs thiocillin-producing strain Bacillus cereus . We show thiocillin variants harboring acid bioorthogonal chemical reactivity can be further modified to create probes...
Nitrogen heterocycles are the key functional and structural elements in both RNA DNA, half a dozen of most important coenzymes, many synthetic drug scaffolds. On other hand, only 3 20 proteinogenic amino acids have nitrogen heterocycles: proline, histidine, tryptophan. This inventory can be augmented microbial proteins by posttranslational modifications downstream leader peptide regions that convert up to 10 serine, threonine, cysteine residues, side chains backbones, into oxazoles,...
Thiazolyl peptides are bacterial secondary metabolites that potently inhibit protein synthesis in Gram-positive bacteria and malarial parasites. Recently, our laboratory others reported this class of trithiazolyl pyridine-containing natural products is derived from ribosomally synthesized preproteins undergo a cascade posttranslational modifications to produce architecturally complex macrocyclic scaffolds. Here, we report the gene cluster responsible for production elongation factor Tu...
The development of immunotherapies for multiple myeloma is critical to provide new treatment strategies combat drug resistance. We report a bispecific antibody against B cell maturation antigen (BiFab-BCMA), which potently and specifically redirects T cells lyse malignant cells. BiFab-BCMA lysed target BCMA-positive lines up 20-fold more than CS1-targeting (BiFab-CS1) developed in an analogous fashion. Further, robustly activated vitro mediated rapid tumor regression orthotopic xenograft...
Berninamycin is a member of the pyridine-containing thiopeptide class antibiotics that undergoes massive posttranslational modifications from ribosomally generated preproteins. has 2-oxazolyl-3-thiazolyl-pyridine core embedded in 35-atom macrocycle rather than typical trithiazolylpyridine cores 26-atom and 29-atom peptide macrocycles. We describe cloning an 11-gene berninamycin cluster Streptomyces bernensis UC 5144, its heterologous expression lividans TK24 venezuelae ATCC 10712, detection...
The thiocillins from Bacillus cereus ATCC 14579 are natural products the broader class of thiazolyl peptides. Their biosynthesis proceeds via extensive post-translational modification a ribosomally encoded precursor peptide. This tailoring involves key step formal cycloaddition between two distal serine residues. In wild-type structure, this forms major macrocycle circumscribed by 26-atoms (shortest path). Results presented herein demonstrate promiscuity last means set "competition"...