Multicellular organisms rely on cell-cell contacts and substance exchange via transmembrane channels such as gap-junctions to maintain their biological behavior. Despite substantial advances in protocell-environment signaling, key challenges persist in developing synthetic channels that mimic the natural gap-junctions for macromolecular transport in protocells. Here, we developed an electrostatic interaction-based strategy for constructing nanoparticle-mediated gap-junction mimics in the membrane of protocells, investigating electrostatic modulation of membrane integrity and subsequent transmembrane signaling regulation. By engineering interactions between charged nanoparticles and protocell membrane, we modulated the contact and higher-order assembly behavior of protocells. We also achieved molecular weight-dependent and concentration gradient-controlled directional transmembrane transport of macromolecular signals between protocells. Further integrating molecular dynamics simulations with experimental validation, we systematically deciphered nanoparticle-membrane interaction dynamics and established the mechanism of electrostatic regulation in transmembrane communication. In summary, this study offers promising insights into the physical principles underlying transmembrane signaling and establishes a foundation for constructing synthetic cellular networks with advanced macromolecular communication functionalities.

The development of nanomedicine that can be efficiently internalized by drug-resistant cancer cells still presents a daunting challenge due to the low uptake capacity caused by various drug resistance-related factors on the cell membrane. Herein, we engineered the surfaces of glycan nanocarriers with negative, neutral, and gradient positive charges, and discovered that positively charged nanocarriers can be anchored onto the cell membrane through electrostatic attraction and thus be efficiently internalized by drug-resistant cancer cells. In contrast, drug-resistant cancer cells do not readily uptake neutral or negatively charged nanocarriers. By proposing a concentric ring fluorescence coefficient (CRFC), we were able to quantify the cell membrane anchoring capabilities of the nanocarriers and found that positively charged nanocarriers have a much stronger anchoring ability toward drug-resistant cell membranes than their neutral and negatively charged counterparts. Interestingly, with the increase of positive charge, the ability of the nanocarriers to become anchored onto cell membranes was further enhanced, thus confirming that electrostatic attraction plays a crucial role in the membrane-anchoring guided cellular uptake. The method of endowing nano-objects with this charge-attracting capability towards negatively charged cell membranes to drive membrane-anchoring mediated cellular uptake illustrates its potential as a universal strategy for engineering nanocarriers to promote the uptake of nanodrugs into drug-resistant cancer cells and thus improve the therapeutic effect.

Conjunctival melanoma (CοΜ) is of high malignancy that diffusely impacts the ocular surface, which is challenging to surgically remove and pose a life-threatening risk. As one of the most suitable treatment options, drug dropping drugs to the eye surface, however, is limited in its clinical application for CoM treatment due to low drug utilization. Using BRAF/MEK inhibitors (BRAF/MEKi) as model drugs, this study proposes a long-lasting eyedrop based on BRAF/MEK dual-targeted liposome-doped dynamic hydrogels with drugs (B/MLDDHs), which can overcome the tear flushing to succeed in effective ocular surface drug delivery. B/MLDDHs encapsulate drugs within lipid bilayers, enhancing drug bioavailability and biocompatibility. The aldehyde dextran and protonated chitosan-based injectable hydrogel system facilitates drug retention on the ocular surface exerting a sustained liposome release, which significantly enhances the drug bioavailability. In therapeutics, this study demonstrates significantly enhanced in vivo therapeutic effect on orthotopic mouse model of CoM. Given their ability to fulfill permeable and long-term release of hydrophobic drugs, B/MLDDHs may serve as a promising platform for non-invasive delivery of various drug molecules, thereby improving therapeutic outcomes for CoM.

Herein we develop a unique differentiated-uptake strategy capable of efficient and high-purity isolation of genuine drug-resistant (DR) cells from three types of drug-surviving cancer cells, which include paclitaxel-surviving human ovarian OVCAR-3 cancer cells and human lung carcinoma A549/Taxol cells, and doxorubicin-surviving human immortalized myelogenous leukemia K562/ADR cells. By using this strategy which relies on fluorescent glycan nanoparticle (FGNP)-based fluorescence-activated cell sorting (FACS) assays, two subpopulations with distinct fluorescences existing in drug-surviving OVCAR-3 cells were separated, and we found that the lower fluorescence (LF) subpopulation consisted of DR cells, while the higher fluorescence (HF) subpopulation was comprised of non-DR cells. Besides, the DR cells and their progenies were found distinct in their increased expression of drug-resistant genes. More intriguingly, by using the FGNP-based FACS assay to detect DR/non-DR phenotypes, we found that the DR phenotype had a potential to differentiate into the non-DR progeny, which demonstrates the differentiation feature of stem-like cancer cells. Further research disclosed that the assay can quantitatively detect the degree of drug resistance in DR cells, as well as the reversal of drug resistance that are tackled by various therapeutic methods. The strategy thus paves the way to develop theranostic approaches associated with chemotherapy-resistance and cancer stemness.