Biology subjects

Wang, L. G.

Publications and source records attributed to Wang, L. G..

4 recordsLinked to original sources

bioRxiv2026

Cyclic immunofluorescence platform using photocleavable linkers for direct antibody labeling enables cancer phenotyping

Advances in spatial proteomics through the development of multiplexed immunostaining platforms have facilitated analyses with increasing cellular and molecular granularity. However, currently available approaches are limited by harsh conditions for signal removal, restricting the number of antigens that can be probed in a single specimen without significant alterations to sample quality and structure. Here we present an approach for direct labeling of primary antibodies with fluorophores using a photocleavable linker (PCL) with a polyethylene glycol spacer (PEG) to enable cyclic immunofluorescence (cyCIF) with gentle signal removal conditions. Our innovative approach uses directly labeled primary antibodies to enhance staining specificity and cyclic immunostaining efficiency, while minimizing nonspecific background signal. Additionally, through integration of the PCL, this approach facilitates gentle cleavage of antibody conjugated fluorophore, preserving sample integrity over multiple rounds of staining. Direct PEG-PCL antibody labeling will promote greater multiplexing by minimizing specimen damage and allow for quantitative analyses of cyCIF spatial data. We demonstrate that cyCIF with PEG-PCL conjugated antibodies can be applied across a variety of cancer subtypes to identify and characterize rare neoplastic cell populations in both tumor tissue and fragile peripheral blood specimens.

biochemistry↗

bioRxiv2026

BBB-Permeable Near-Infrared Oxazine Fluorophores for White Matter Tract Imaging

Preserving critical functional tissue remains a major challenge in fluorescence-guided surgery (FGS), particularly in neurosurgery where injury to white matter tracts can lead to lasting neurological deficits. Probes for preventing iatrogenic injury of the peripheral nervous system have highlighted the potential of real-time intraoperative guidance for nerve-sparing surgery, supporting improved preservation of critical functional tissue, but analogous tools for the central nervous system remain limited. While there are some awake mapping and MRI based techniques that try to minimize the resection of white matter tracts (WMTs), they do not provide surgeons with real-time visualization. To address this gap, we rationally translated a peripheral nerve targeted oxazine fluorophore platform into a library of novel oxazine dyes that can penetrate the blood-brain barrier (BBB), exhibit specificity for white matter tissue and maintain spectral compatibility with clinically relevant imaging systems. Guided by our fluorophore medicinal chemistry platform, this work includes library design, photophysical property characterization, in vivo WMTs specificity screening, and in vivo imaging window assessment of the lead candidates to evaluate neurosurgical workflow compatibility. The lead candidate identified from this library demonstrated strong in vivo WMTs-specific fluorescence intensity, contrast and imaging window compatible with neurosurgical workflows, establishing BBB-permeable oxazine fluorophores as a promising scaffold for future structure-sparing neurosurgical FGS.

bioengineering↗

bioRxiv2025

Engineering NIR probes to enhance affinity and clinical workflow compatibility for prostate cancer imaging

Positive surgical margins following radical prostatectomy increase the risk of biochemical recurrence and subsequent disease progression. Fluorescence guided surgery (FGS) using targeted contrast agents has shown clinical benefits for several cancer types. However, current prostate cancer targeted imaging probes exhibit long pharmacokinetic (PK) profiles, necessitating extended waiting periods or repeated hospital visits, limiting their integration into standard clinical workflow. To overcome this critical clinical compatibility challenge, we developed an innovative tri-compartment, chemistry-driven probe design strategy. Specifically, we developed a congeneric library of near infrared (NIR) water soluble fluorescent probes incorporating: (1) a glutamic acid-urea-lysine (EuK) ligand targeting prostate specific membrane antigen (PSMA); (2) a NIR heptamethine cyanine fluorophore optimized for enhanced PSMA binding via secondary binding sites interactions; and (3) distinct PK modulators residing outside the PSMA binding pocket to promote rapid off-target tissue clearance. While molecular docking scores, photophysical properties and live-cell staining results showed similar overall performance, probes bearing PK modulators produced stronger tumor-specific fluorescence in vivo than the control lacking a PK modulator. This effort enabled identification of a lead probe with robust tumor targeting and accelerated off-target clearance, providing optimal tumor-specific signal and contrast in a timeframe, fully compatible with robotic-assisted radical prostatectomy (RARP) timelines.

pharmacology and toxicology↗

bioRxiv2020

Crosstalk between invadopodia and the extracellular matrix

The scaffold protein Tks5 is required for invadopodia-mediated cancer invasion both in vitro and in vivo. We have previously also revealed a role for Tks5 in tumor cell growth using three-dimensional (3D) culture model systems and mouse transplantation experiments. Here we use both 3D and high-density fibrillar collagen (HDFC) culture to demonstrate that native type I collagen, but not a form lacking the telopeptides, stimulated Tks5-dependent growth, which was dependent on the DDR collagen receptors. We used microenvironmental microarray (MEMA) technology to determine that laminin, collagen I, fibronectin and tropoelastin also stimulated invadopodia formation. A Tks5-specific monoclonal antibody revealed its expression both on microtubules and at invadopodia. High- and super-resolution microscopy of cells in and on collagen was then used to place Tks5 at the base of invadopodia, separated from much of the actin and cortactin, but coincident with both matrix metalloprotease and cathepsin proteolytic activity. Inhibition of the Src family kinases, cathepsins or metalloproteases all reduced invadopodia length but each had distinct effects on Tks5 localization. These studies highlight the crosstalk between invadopodia and extracellular matrix components, and reveal the invadopodium to be a spatially complex structure.

cancer biology↗