<i>In Situ</i> Visualization of PD-L1-Specific Glycosylation on Tissue Sections

Huang, Mengjiao; Zhu, Lin; Kang, Siyin; Chen, Fude; Wei, Xinyu; Lin, Liang; Chen, Xiaofeng; Wang, Wei; Zhu, Zhi; Yang, Chaoyong; Song, Yanling

doi:10.1021/acs.analchem.1c03287

Cited by 22 publications

(23 citation statements)

References 31 publications

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“…First, we designed a circular DNA template containing sequences complementary to PDL1 aptamer, CpG oligonucleotide (ODN), and G-quadruplex functional sequences for RCA reaction (Table S1). Specifically, the PDL1 aptamer can bind to PDL1 on the tumor cell surface, blocking the PD1-PDL1 interaction. ,, The CpG ODNs conjugate to the Toll-like acceptors to trigger robust immune stimulation and reinforce inhibition of tumor cells. ,,, The G-quadruplex is used to intercalate the porphyrin photosensitizer TMPyP 4 to achieve high drug loading capacity . AptPDL1/CpG@DNF-TMPyP 4 (PCT) was obtained by self-assembly of the long single-stranded DNA (ssDNA) generated from RCA reaction (Scheme a).…”

Section: Resultsmentioning

confidence: 99%

Photocontrolled Spatiotemporal Delivery of DNA Immunomodulators for Enhancing Membrane-Targeted Tumor Photodynamic Immunotherapy

Wang

Liu

Duan

et al. 2022

ACS Appl. Mater. Interfaces

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Immunotherapy is emerging as a paradigm-shifting modality for treatment cancer. However, systemic administration of immunomodulators is usually accompanied by extra-tumor toxicity and adverse immune effects. Precise delivery of immunomodulators with a highly controllable system may provide a solution for this issue. Here, we developed a photocontrolled DNA nanomedicine for localized delivery of DNA immunomodulators to enhance membrane-targeted photodynamic immunotherapy. Specifically, the DNA nanomedicine is composed of long tandemly repeated functional DNA sequences (PDL1 aptamers and CpG) with a photosensitizer (TMPyP4) inserted into the DNA structure, providing high drug-loading capacity. By blocking the surface PDL1 aptamer with a pHLIP-modified cDNA, the DNA nanomedicine does not induce any obvious immune response and can be specifically delivered and anchored to the tumor membrane. Under localized irradiation, photodynamically generated reactive oxygen species (ROS) causes breakage of DNA sequences, which triggers the collapse of the nanostructure and release of internal DNA immunomodulators. Under localized illumination, photodynamically generated ROS can cause DNA sequence breaks, triggering the collapse of nanostructures and the release of internal DNA immunomodulators thus enhancing membrane-targeted photodynamic immunotherapy. We have demonstrated that the developed DNA nanomedicine can drive efficient immune responses in tumor tissue without perceptibly interfering off-tumor immunity, resulting in efficient antitumor treatment while mitigating systemic toxicity.

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Section: Resultsmentioning

confidence: 99%

Photocontrolled Spatiotemporal Delivery of DNA Immunomodulators for Enhancing Membrane-Targeted Tumor Photodynamic Immunotherapy

Wang

Liu

Duan

et al. 2022

ACS Appl. Mater. Interfaces

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“…As a proof of concept, glycosylation of exosomal programmed cell death ligand 1 (exoPD-L1), which has been proven to play a pivotal role in regulating immune evasion [34], was selected as a model. The PD-L1 aptamer MJ5C was selected to design Aptamer-T2 probe, because MJ5C has a high binding affinity against glycosylated PD-L1 and its binding does not affect the interaction of PD-L1 and PD-1 [27,28]. Other corresponding probes (DBCO-T1, connector, H1, and H2) were also constructed for proximity-induced HCR assembly (Table S1, Supporting Information online).…”

Section: Validation Of Hcr Assembly In Buffermentioning

confidence: 99%

“…This strategy provides several significant advantages. Firstly, compared with antibody labelling, the much smaller aptamer enables the recognition with minimum interference on the natural function of the target protein [27,28]. Moreover, aptamers selected through in vitro evolution can lead to a series of aptamer sequences binding different epitopes of target protein to provide recognition elements targeting nonfunctional epitopes of the target glycoprotein [29,30].…”

Section: Introductionmentioning

confidence: 99%

Amplified visualization and function exploration of exosomal protein-specific glycosylation using hybridization chain reaction from non-functional epitope

et al. 2022

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Exosomal glycoproteins play significant roles in many physiological and pathological procedures. However, the current methods for studying exosomal glycoproteins have low sensitivity or can affect exosomal biological function. Herein, we developed a proximity dual-tagging strategy using an induced hybridization chain reaction (HCR) from the target's non-functional epitope for amplified visualization and functional exploration of exosomal protein-specific glycosylation. This strategy leverages dualtagging based on the aptamer with little influence on target function and metabolic glycan labelling, and the rigid product and high sensitivity of HCR. The method improves the signal of visualizing exosomal PD-L1 (exoPD-L1) by 7.7-fold compared with the signal without HCR amplification without affecting the natural exoPD-L1/PD-1 interaction. As a result, we verified that the interaction between exoPD-L1 and PD-1 positive cells is positively correlated to the glycosylation level of exoPD-L1. Overall, we have developed a sensitive method with little functional influence to visualize exosomal protein-specific glycosylation in situ, offering a powerful tool for studying the biological implications of exosomal glycoproteins.

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“… 30,32,33 This process plays an important role in cell growth, differentiation, transformation, adhesion and tmour immune monitoring 34,35 . Researchers used a FLAG strategy for in situ visualization of protein‐specific glycosylation on tissue slices, this strategy has been used to visualize the states and distribution of PD‐L1‐specific glycosylation in lung cancer tissue biopsies 36 . Glycosylation of co‐suppressive molecules (i.e.…”

Section: Source Structure and Glycosylation Of Pd‐l1mentioning

confidence: 99%

“…34,35 Researchers used a FLAG strategy for in situ visualization of protein-specific glycosylation on tissue slices, this strategy has been used to visualize the states and distribution of PD-L1-specific glycosylation in lung cancer tissue biopsies. 36 Glycosylation of co-suppressive molecules (i.e. molecules with a major biological function involved in suppressing immune responses) including PD-L1 is critical for the regulation of immunosuppression and immune surveillance.…”

Section: Source Structure and Glycosylation Of Pd -L1mentioning

confidence: 99%

Research progress of PD‐L1 non‐glycosylation in cancer immunotherapy

et al. 2022

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Cancer is the second leading cause of death among human diseases.Immunotherapy has opened a new chapter in cancer treatment. However, the emergence of immune escape mechanisms severely limited its application. The high expression of PD-L1 on tumour cells is an important means for cancer cells to achieve immune escape via binding to PD-1 on immune cells. Recent studies have shown that PD-L1 expression in most cancer cells is over-glycosylated.Glycosylation is a fundamental and extensive post-translational modification of eukaryotic membrane-binding proteins, which affects a variety of biological activities, including protein folding, solubility, stability and ligand-receptor interactions. PD-L1 glycosylation initiates in the endoplasmic reticulum (ER) and completes in the Golgi apparatus, which has a complex relationship with cancer development. Importantly, non-glycosylated PD-L1 attenuates protein stability and PD-1 interactions. These processes are essential regulatory mechanisms that modulate immunosuppression and immune surveillance in cancer patients.Therefore, non-glycosylated PD-L1 may be a potentially promising strategy to improve the efficacy of checkpoint blockade therapy for cancer. In this review, we have described the PD-L1 glycosylation processes and provided evidence for the role of non-glycosylated PD-L1 in anti-tumour-related signalling pathways. Furthermore, strategies for non-glycosylation of PD-L1 using small molecule inhibitors, gene therapy and various enzymes in the synthetic glycosylation pathway are discussed. Finally, the detection of PD-L1 expression is enhanced by its deglycosylation, and we have summarized the development, application and clinical application potential of antibody therapies targeting PD-1/PD-L1 glycosylation.

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In Situ Visualization of PD-L1-Specific Glycosylation on Tissue Sections

Abstract: Scheme 1. Working Principle of the FLAG Strategy a a This FLAG strategy offers visualization of PD-L1-specific glycosylation on tissue sections through FRET between the PD-L1 aptamer (MJ5C)-labeled FAM and a lectin (Con A)-conjugated Cy3.

Cited by 22 publications

References 31 publications

Photocontrolled Spatiotemporal Delivery of DNA Immunomodulators for Enhancing Membrane-Targeted Tumor Photodynamic Immunotherapy

Photocontrolled Spatiotemporal Delivery of DNA Immunomodulators for Enhancing Membrane-Targeted Tumor Photodynamic Immunotherapy

Amplified visualization and function exploration of exosomal protein-specific glycosylation using hybridization chain reaction from non-functional epitope

Research progress of PD‐L1 non‐glycosylation in cancer immunotherapy

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