Biology subjects

Li, J. P.

Publications and source records attributed to Li, J. P..

5 recordsLinked to original sources

bioRxiv2025

Cocktail Chemical Labeling for In-Depth Surfaceome Profiling of Bone-Marrow Derived Dendritic Cells

Cell surface proteins (CSPs) are crucial identifiers for cell types and states, especially in dendritic cells (DCs). Current proteomic methods for profiling CSPs are limited by their hydrophobic nature and low abundance, which often require genetic or cell surface engineering and exhibit biased and insufficient labeling efficiency. Herein, we report [Ru(bpy)3]Cl2 (Ru) for effective biotinylation on the cell surfaceome via a simple "mix and lighten" method. The versatile photoredox pathways of Ru are leveraged using a probe cocktail of biotin-phenol and biotin-hydrazide for improved substrate coverage. The "cocktail" labeling strategy results in reproducible identification of up to 733 plasma membrane proteins on HeLa cells, and is further applied to map dynamic changes in the surfaceome during the differentiation of primary bone marrow-derived dendritic cells, which demonstrates a user-friendly and deep-surfaceome-coverage tool for profiling dynamic changes in primary cells, with potential implications for cell identities, functional states, and novel drug targets.

cell biology↗

bioRxiv2023

Anti-PD-1-iRGD Peptide Conjugate Boosts Antitumor Efficacy via Engagement Augmentation and Penetration Enhancement of T cells

Despite the important breakthroughs of immune-checkpoint inhibitors (ICIs) in recent years, the overall objective response rate (ORR) remains limited in various cancers. Here, we synthesized programmed cell death protein-1 (PD-1) antibody iRGD conjugate (PD-1-(iRGD)2) through glycoengineering and bio-orthogonal reaction. PD-1-(iRGD)2 exhibited extra iRGD receptor dependent affinity to several cancer cell lines rather than normal cell lines. Via dual targeting, PD-1-(iRGD)2 engageed tumor cells and T cells thus mediating T cell activation and facilitating tumor elimination. Besides, the attachment of iRGD impressively improved the penetrability of both PD-1 antibody and PD-1+ T cells. In multiple syngeneic mouse models, PD-1-(iRGD)2 effectively reduced tumor growth with satisfactory biosafety. Moreover, results of flow cytometry and single-cell RNA-seq revealed that PD-1-(iRGD)2 remodeled the tumor microenvironment (TME) and expanded a unique population of "better effector" CD8+ tumor infiltrating T cells (TILs) expressing stem and memory associated genes including Tcf7, Il7r, Lef1 and Bach2. Conclusively, PD-1-(iRGD)2 could be a novel and promising therapeutic approach for cancer immunotherapy. Statement of significanceDesigned against the clinical dilemma of unsatisfied response rate after contemporary cancer immunotherapy, PD-1-(iRGD)2 engages T cells and tumor cells, promotes T cell infiltration and expands a unique population of "better effectors" with enhanced therapeutic potential for the treatment of cancer.

immunology↗

bioRxiv2022

Reducing the Complexity of Fc Glycan Enables the Construction of Antibody Conjugates with Unexpected High Efficiency and Payload Capacity via Glycoengineering

Glycoengineering has been demonstrated to be a powerful tool to construct site-specific antibody conjugates. However, with most glycoengineering strategies, several hours to days are needed to complete the reaction, and the payloads are limited to small molecules. Herein, we show that reducing the complexity of Fc glycan could dramatically boost the enzymatic glycoengineering of IgG molecules to achieve unexpectedly high efficiency and payload capacity. Notably, antibodies with the modified Fc glycan were successfully conjugated with biomacromolecules, e.g., oligonucleotides and nanobodies, in one step by an engineered fucosyltransferase. Moreover, the conversion of this chemoenzymatic engineering could be finished in minutes, or up to hours, affording homogeneous products. The trisaccharide linker of these antibody conjugates exhibits excellent hydrophilicity and stability and reduced ADCC activity. Using this platform, we successfully synthesized an antibody-conjugate-based HER2/CD3 bispecific antibody and applied it to selectively destroy patient-derived cancer organoids by reactivating endogenous T cells inside the organoid. These results suggest that this bispecific antibody format has a high translational value.

biochemistry↗

bioRxiv2022

Chemoenzymatic Measurement of Cell-surface Glycan in Single-cell Multiomics: LacNAc as an Example

Despite the rich information of a cells physiological state encoded in the dynamic changes of cell-surface glycans, methods of capturing glycosylation states at the single-cell level are quite limited. Here we report a chemoenzymatic single-cell N-acetyllactosamine (LacNAc) detection method via tagging the LacNAc with a specific DNA barcode. Compared to the lectin-based glycan detection, the chemoenzymatic labeling does not change the transcriptional status of immune cells and is more compatible with scRNA-seq. Integrated analysis of LacNAc and transcriptome of T cells at a single-cell level reveals that the quantity of cell-surface LacNAc is significantly upregulated in activated CD8+ T cells but maintained at the basal level in quiescent CD8+ T cells (i.e., naive and central memory T cells). Further analysis confirms that the LacNAc level is positively correlated to the glycolytic activity of CD8+ T cells at all statues. Taken together, our study demonstrates the feasibility of chemoenzymatic detection of cell-surface glycan in single-cell RNA sequencing-based multiomics with information of TCR sequence and cell-surface epitopes (i.e., scTCR and CITE-seq) and offers a new way to characterize the biological role of glycan in diversified physiological states.

immunology↗

bioRxiv2020

Detecting Tumor Specific Antigen-Reactive T cells from Tumor Infiltrating Lymphocytes via Interaction Dependent Fucosyl-biotinylation

Re-activation and clonal expansion of tumor specific antigen (TSA)-reactive T cells are critical to the success of checkpoint blockade and adoptive transfer of tumor-infiltrating lymphocyte (TIL) based therapies. There are no reliable markers to specifically identify the repertoire of TSA-reactive T cells due to their heterogeneous composition. Here we introduce FucoID as a general platform to detect endogenous antigen-specific T cells and study their biology. Through this interaction dependent labeling approach, TSA-reactive T cells can be detected and separated from bystander T cells in primary tumor digests based on their cell-surface enzymatic fucosyl-biotinylation. Compared to bystander TILs, TSA-reactive TILs possess a distinct TCR repertoire and unique gene features. Though exhibiting a dysfunctional phenotype, this subset of TILs possesses substantial capabilities of proliferation and tumor specific killing. FucoID features genetic manipulation-free procedures and a quick turnover cycle, and therefore should have the potential of accelerating the pace of personalized cancer treatment. HighlightsInteraction dependent fucosylation enables the detection and isolation of bona fide intratumoral tumor specific antigen-reactive T cells Tumor specific antigen-reactive CD8+ T cells possess capabilities to be expanded and adoptively transferred for tumor control Tumor specific antigen-reactive CD8+ T cells feature oligoclonal expansion and upregulate genes for the steroid biosynthesis and metabolic process Intratumoral bystander CD8+ T cells can be separated into two groups based on PD-1 expression that feature distinct gene modules

immunology↗