Publications

The development of effective antibody therapeutics has been hampered by a lack of methods to measure drug delivery and activity within tumors at single-cell resolution. Here we introduce single-cell spatial pharmacobiology (SSP), an experimental and analytical framework that integrates in situ imaging of a systemically infused, fluorescently labeled therapeutic antibody with high-plex spatial proteomics to quantify antibody distribution, target engagement and tumor microenvironment (TME) architecture. We applied SSP to tumor tissues from participants with head and neck squamous cell carcinoma and pancreatic ductal adenocarcinoma who received the antibody panitumumab-IRDye800 in phase 1 trials. SSP identified pronounced spatial heterogeneity in single-cell drug delivery and target engagement, shaped by conserved stromal barriers, including periostin-rich extracellular matrix assemblies and fibroblast-activation-protein-positive cancer-associated fibroblast neighborhoods, which were associated with reduced antibody delivery in both tumor types. SSP measures drug-target-TME interactions in human tumors and can support studies of resistance mechanisms, dosing strategies and discovery of spatial biomarkers for precision oncology.

JTD Keywords: Cancer, Diffusion, Fibroblasts, Gemcitabine, Head, Inhibitor, Monoclonal-antibodies, Multiplex, Panitumumab-irdye800cw, Transport

Skin is a barrier that protects us against physical, chemical and biological agents. However, any damage to the skin can disrupt this barrier and therefore compromise its function leading to sometimes catastrophic consequences like sepsis. Thus, methods to detect early signs of infection are necessary. In this work, we have developed a straightforward method for producing 2D nanomembranes with regularly spaced 1D metallic nanostructures integrating sensing capabilities to pH and NADH (nicotinamide adenine dinucleotide), which are critical analytes revealing infection. To achieve this, we have successfully fabricated a bilayered nanomembrane combining a pH-responsive polyaniline (PANI) layer and a nanoperforated poly(lactic acid) (PLA) layer containing gold nanowires (Au NWs) as NADH sensing element. SEM, FTIR, Raman and AFM techniques revealed the formation of the bilayered PANI/PLA nanomembrane and the successful incorporation of the Au NWs inside the nanoperforations. The resulting bilayered nanomembrane showed significant flexibility and conformability onto different substrates due to the softness of the polymers and the ultrathin thickness with stiffness values similar to human skin. These nanomembranes also exhibited remarkable electrochemical sensing performance towards pH and NADH detection. Thus, the nanomembrane displayed linearity with good sensitivity (47 mVpH-1) in the critical pH range 4-10 and fast response time (10 s). On the other hand, PANI/PLA-Au nanomembranes also allowed the quantitative sensing of NADH with a limit of detection of 0.39 mM and a sensitivity of 1 mu A cm-2 mM-1 in the concentration range 0-5 mM.

JTD Keywords: Bilayered nanomembranes, Biomimetic membranes, Free-standing films, Gold nanowires, Nad, Nanoparticles, Pani, Ph and nadh sensor, Pla, Polyaniline