Dan Yang

Biomacromolecules in physiological environments would adsorb onto the nanoparticles (NPs) to form corona layers, in which protein coronas (PCs) are the major constituent. PCs always play diverse influences on the fate of NPs in vitro and in vivo, especially for active-targeting NPs (e.g., transferrin-modified nanoparticles, Tf-NP). In order to eliminate the inhibition of PCs on the efficiency of Tf-NP, the precoated Tf-NP with bovine serum albumin (BSA, B@Tf-NP) was designed to fabricate an "active PCs" (PCs formed by artificial modification) against the "passive PCs" (PCs formed in the biological environments), which was inspired by the formation pattern of PCs. The results indicated that B@Tf-NP had similar particle size, dispersion, and physical stability with Tf-NP. Surprisingly, B@Tf-NP enhanced the cellular uptake in enterocytes and permeability in intestinal tract of mice. Notably, the concentration ratio of BSA to Tf that could ensure Tf revealed timely during the interacted process was considered to be appropriate. These findings provide an easy while efficient design platform for active-targeting NPs to overcome the biomacromolecular barrier in oral administration.

The epithelial mucosa is a key biological barrier faced by gastrointestinal, intraoral, intranasal, ocular, and vaginal drug delivery. Ligand-modified nanoparticles demonstrate excellent ability on this process, but their efficacy is diminished by the formation of protein coronas (PCs) when they interact with biological matrices. PCs are broadly implicated in affecting the fate of NPs in vivo and in vitro, yet few studies have investigated PCs formed during interactions of NPs with the epithelial mucosa, especially mucus. In this study, we constructed transferrin modified NPs (Tf-NPs) as a model and explored the mechanisms and effects that epithelial mucosa had on PCs formation and the subsequent impact on the transcellular transport of Tf-NPs. In mucus-secreting cells, Tf-NPs adsorbed more proteins from the mucus layers, which masked, displaced, and dampened the active targeting effects of Tf-NPs, thereby weakening endocytosis and transcellular transport efficiencies. In mucus-free cells, Tf-NPs adsorbed more proteins during intracellular trafficking, which enhanced transcytosis related functions. Inspired by soft coronas and artificial biomimetic membranes, we used mucin as an "active PC" to precoat Tf-NPs (M@Tf-NPs), which limited the negative impacts of "passive PCs" formed during interface with the epithelial mucosa and improved favorable routes of endocytosis. M@Tf-NPs adsorbed more proteins associated with endoplasmic reticulum-Golgi functions, prompting enhanced intracellular transport and exocytosis. In summary, mucus shielded against the absorption of Tf-NPs, but also could be employed as a spear to break through the epithelial mucosa barrier. These findings offer a theoretical foundation and design platform to enhance the efficiency of oral-administered nanomedicines.

Cancer immunotherapy is expected to achieve tumor treatment mainly by stimulating the patient's own immune system to kill tumor cells. However, the low immunogenicity of the tumor and the poor efficiency of tumor antigen presentation result in a variety of solid tumors that do not respond to immunotherapy. Herein, we designed a proton-gradient-driven porphyrin-based liposome (PBL) with highly efficient Toll-like receptor 7 (TLR7) agonist (imiquimod, R837) encapsulation (R837@PBL). R837@PBL rapidly released R837 in the acid microenvironment to activate the TLR in the endosome inner membrane to promote bone-marrow-derived dendritic cell maturation and enhance antigen presentation. R837@PBL upon laser irradiation triggered immunogenic cell death of tumor cells and tumor-associated antigen release after subcutaneous injection, activated TLR7, formed in situ tumor nanoadjuvants, and enhanced the antigen presentation efficiency. Photoimmunotherapy promoted the infiltration of cytotoxic T lymphocytes into tumor tissues, inhibited the growth of the treated and abscopal tumors, and exerted highly effective photoimmunotherapeutic effects. Hence, our designed in situ tumor nanoadjuvants are expected to be an effective treatment for treated and abscopal tumors, providing a novel approach for synergistic photoimmunotherapy of tumors.

Cancer immunotherapy, such as immune checkpoint blockade, chimeric antigen receptor, and cytokine therapy, has emerged as a robust therapeutic strategy activating the host immune system to inhibit primary and metastatic lesions. However, low tumor immunogenicity (LTI) and immunosuppressive tumor microenvironment (ITM) severely compromise the killing effect of immune cells on tumor cells, which fail to evoke a strong and effective immune response. As an exogenous stimulation therapy, phototherapy can induce immunogenic cell death (ICD), enhancing the therapeutic effect of tumor immunotherapy. However, the lack of tumor targeting and the occurrence of immune escape significantly reduce its efficacy in vivo, thus limiting its clinical application. Nanophotoimmunotherapy (nano-PIT) is a precision-targeted tumor treatment that co-loaded phototherapeutic agents and various immunotherapeutic agents by specifically targeted nanoparticles (NPs) to improve the effectiveness of phototherapy, reduce its phototoxicity, enhance tumor immunogenicity, and reverse the ITM. This review will focus on the theme of nano-PIT, introduce the current research status of nano-PIT on converting "cold" tumors to "hot" tumors to improve immune efficacy according to the classification of immunotherapy targets, and discuss the challenges, opportunities, and prospects.

Numerous literatures have shown the advantages of Pickering emulsion (PE) for the delivery of bioactive ingredients in the fields of food, medicine, and cosmetics, among others. On this basis, the multi‐loading mode of bioactives (internal phase encapsulation and/or loading at the interface) in small molecular bioactives nanocrystal‐stabilized PE (BNC‐PE) enables them higher loading efficiencies, controlled release, and synergistic or superimposed effects. Therefore, BNC‐PE offers an efficacious delivery system. In this review, we briefly summarize BNC‐PE fabrication and characterization, with a focus on the processes of possible evolution and absorption of differentially applied BNC‐PE when interacting with the body. In addition, methods of monitoring changes and absorption of BNC‐PE in vivo, from the nanomaterial perspective, are also introduced. The purpose of this review is to provide an accessible and comprehensive methodology for the characterization and evaluation of BNC‐PE after formulation and preparation, especially in relation to biological assessment and detailed mechanisms throughout the absorption process of BNC‐PE in vivo.

A tremendous number of proteins participate in the delivery and transport process of nanomedicines. Nanoprotein interactions not only mediate drug delivery but also determine drug safety. In the field of biomedical sciences, the epithelial barrier is a huge challenge for gastrointestinal, intratracheal, intranasal, vaginal, and intrauterine delivery of nanomedicines. However, the molecular mechanisms by which nanomedicines cross tissue or cell barriers are not well understood. Here, we explored the nanoprotein interactions during the transcytosis of nanoparticles across the epithelial barrier by focusing on the transport pathway and mechanisms. Due to the limitations of traditional methods in resolving nanoprotein interactions, we developed a backward analysis strategy. By simultaneously analyzing the protein corona on the particle surface and the cellular response after transcytosis, we integrated the information on both directly and indirectly interacting proteins, establishing a holistic nanoprotein interaction atlas. It revealed the dominant role of the EV/ER/Golgi/SV pathway in the transcytosis of nanoparticles. More importantly, based on the established atlas, we discovered the association of Wnt/β-catenin signaling with nanoparticle transportation. The endocytosis for entering cells and exocytosis/transcytosis for leaving cells were differently regulated by the Wnt pathway. Notably, this regulatory effect was dependent on the particle size. Bigger nanoparticles departed from cells through the exocytosis pathway faster because of the specific bridging effect on the Wnt-Frizzled interaction and the feedback loop construction based on the exosomes. This mechanism gives an interpretation at the molecular level to the transcytosis dilemma of larger nanoparticles. Moreover, the size-dependent Wnt/β-catenin signaling pathway provides a promising regulatory and screening platform for the transportation of different nanomedicines through the epithelial barrier.