参考文献

Interpreting how multicellular interactions in the tumor affect resistance pathways to BRAF and MEK1/2 MAPK inhibitors (MAPKi) remains a challenge. To investigate this,we profiled global ligand-receptor interactions among tumor and stromal/immune cells from biopsies of MAPK-driven disease. MAPKi increased tumor-associated macrophages (TAMs) in some patients,which correlated with poor clinical response,and MAPKi coamplified bidirectional tumor-TAM signaling via receptor tyrosine kinases (RTKs) including AXL,MERTK,and their ligand GAS6. In xenograft tumors,intravital microscopy simultaneously monitored in situ single-cell activities of multiple kinases downstream of RTKs,revealing MAPKi increased TAMs and enhanced bypass signaling in TAM-proximal tumor cells. As a proof-of-principle strategy to block this signaling,we developed a multi-RTK kinase inhibitor nanoformulation that accumulated in TAMs and delayed disease progression. Thus,bypass signaling can reciprocally amplify across nearby cell types,offering new opportunities for therapeutic design. View Publication

Increasing occurrence of moderate to severe intrauterine adhesion (IUA) is seriously affecting the quality of human life. The aim of the study was to establish IUA models in nonhuman primates and to explore the dual repair effects of human umbilical cord-derived mesenchymal stem cells (huMSCs) loaded on autocrosslinked hyaluronic acid gel (HA-GEL) on endometrial damage and adhesion. Here,we recorded the menstrual cycle data in detail with uterine cavities observed and endometrial tissues detected after intervention,and the thicker endometria,decreased amount of fibrotic formation,increased number of endometrium glands,etc.,suggested that both HA-GEL and huMSC/HA-GEL complexes could partially repair IUA caused by mechanical injury,but huMSC/HA-GEL complex transplantation had notable dual repair effects: a reliable antiadhesion property and the promotion of endometrial regeneration. View Publication

Our understanding of the signalling pathways regulating early human development is limited,despite their fundamental biological importance. Here,we mine transcriptomics datasets to investigate signalling in the human embryo and identify expression for the insulin and insulin growth factor 1 (IGF1) receptors,along with IGF1 ligand. Consequently,we generate a minimal chemically-defined culture medium in which IGF1 together with Activin maintain self-renewal in the absence of fibroblast growth factor (FGF) signalling. Under these conditions,we derive several pluripotent stem cell lines that express pluripotency-associated genes,retain high viability and a normal karyotype,and can be genetically modified or differentiated into multiple cell lineages. We also identify active phosphoinositide 3-kinase (PI3K)/AKT/mTOR signalling in early human embryos,and in both primed and na{\{i}}ve pluripotent culture conditions. This demonstrates that signalling insights from human blastocysts can be used to define culture conditions that more closely recapitulate the embryonic niche." View Publication

Self-assembling virus-like particles represent highly attractive tools for developing next-generation vaccines and protein therapeutics. We created ADDomer,an adenovirus-derived multimeric protein-based self-assembling nanoparticle scaffold engineered to facilitate plug-and-play display of multiple immunogenic epitopes from pathogens. We used cryo–electron microscopy at near-atomic resolution and implemented novel,cost-effective,high-performance cloud computing to reveal architectural features in unprecedented detail. We analyzed ADDomer interaction with components of the immune system and developed a promising first-in-kind ADDomer-based vaccine candidate to combat emerging Chikungunya infectious disease,exemplifying the potential of our approach. View Publication

Patient-derived organoids (PDOs) have recently emerged as robust preclinical models; however,their potential to predict clinical outcomes in patients has remained unclear. We report on a living biobank of PDOs from metastatic,heavily pretreated colorectal and gastroesophageal cancer patients recruited in phase 1/2 clinical trials. Phenotypic and genotypic profiling of PDOs showed a high degree of similarity to the original patient tumors. Molecular profiling of tumor organoids was matched to drug-screening results,suggesting that PDOs could complement existing approaches in defining cancer vulnerabilities and improving treatment responses.We compared responses to anticancer agents ex vivo in organoids and PDO-based orthotopic mouse tumor xenograft models with the responses of the patients in clinical trials. Our data suggest that PDOs can recapitulate patient responses in the clinic and could be implemented in personalized medicine programs. View Publication

High-grade gliomas are lethal brain cancers whose progression is robustly regulated by neuronal activity. Activity-regulated release of growth factors promotes glioma growth,but this alone is insufficient to explain the effect that neuronal activity exerts on glioma progression. Here we show that neuron and glioma interactions include electrochemical communication through bona fide AMPA receptor-dependent neuron-glioma synapses. Neuronal activity also evokes non-synaptic activity-dependent potassium currents that are amplified by gap junction-mediated tumour interconnections,forming an electrically coupled network. Depolarization of glioma membranes assessed by in vivo optogenetics promotes proliferation,whereas pharmacologically or genetically blocking electrochemical signalling inhibits the growth of glioma xenografts and extends mouse survival. Emphasizing the positive feedback mechanisms by which gliomas increase neuronal excitability and thus activity-regulated glioma growth,human intraoperative electrocorticography demonstrates increased cortical excitability in the glioma-infiltrated brain. Together,these findings indicate that synaptic and electrical integration into neural circuits promotes glioma progression. View Publication

Vaccine efficacy can be increased by arraying immunogens in multivalent form on virus-like nanoparticles to enhance B-cell activation. However,the effects of antigen copy number,spacing and affinity,as well as the dimensionality and rigidity of scaffold presentation on B-cell activation remain poorly understood. Here,we display the clinical vaccine immunogen eOD-GT8,an engineered outer domain of the HIV-1 glycoprotein-120,on DNA origami nanoparticles to systematically interrogate the impact of these nanoscale parameters on B-cell activation in vitro. We find that B-cell signalling is maximized by as few as five antigens maximally spaced on the surface of a 40-nm viral-like nanoparticle. Increasing antigen spacing up to {\~{}}25-30 nm monotonically increases B-cell receptor activation. Moreover,scaffold rigidity is essential for robust B-cell triggering. These results reveal molecular vaccine design principles that may be used to drive functional B-cell responses. View Publication

Although the association between the microbiome and IBD and liver diseases is known,the cause and effect remain elusive. By connecting human microphysiological systems of the gut,liver,and circulating Treg and Th17 cells,we created a multi-organ model of ulcerative colitis (UC) ex vivo. The approach shows microbiome-derived short-chain fatty acids (SCFAs) to either improve or worsen UC severity,depending on the involvement of effector CD4 T cells. Using multiomics,we found SCFAs increased production of ketone bodies,glycolysis,and lipogenesis,while markedly reducing innate immune activation of the UC gut. However,during acute T cell-mediated inflammation,SCFAs exacerbated CD4+ T cell-effector function,partially through metabolic reprograming,leading to gut barrier disruption and hepatic injury. These paradoxical findings underscore the emerging utility of human physiomimetic technology in combination with systems immunology to study causality and the fundamental entanglement of immunity,metabolism,and tissue homeostasis. View Publication

Our previous study demonstrated that conditional reprogramming (CR) allows the establishment of patient-derived normal and tumor epithelial cell cultures from a variety of tissue types including breast,lung,colon and prostate. Using CR,we have established matched normal and tumor cultures,GUMC-29 and GUMC-30 respectively,from a patient's prostatectomy specimen. These CR cells proliferate indefinitely in vitro and retain stable karyotypes. Most importantly,only tumor-derived CR cells (GUMC-30) produced tumors in xenografted SCID mice,demonstrating maintenance of the critical tumor phenotype. Characterization of cells with DNA fingerprinting demonstrated identical patterns in normal and tumor CR cells as well as in xenografted tumors. By flow cytometry,both normal and tumor CR cells expressed basal,luminal,and stem cell markers,with the majority of the normal and tumor CR cells expressing prostate basal cell markers,CD44 and Trop2,as well as luminal marker,CD13,suggesting a transit-amplifying phenotype. Consistent with this phenotype,real time RT-PCR analyses demonstrated that CR cells predominantly expressed high levels of basal cell markers (KRT5,KRT14 and p63),and low levels of luminal markers. When the CR tumor cells were injected into SCID mice,the expression of luminal markers (AR,NKX3.1) increased significantly,while basal cell markers dramatically decreased. These data suggest that CR cells maintain high levels of proliferation and low levels of differentiation in the presence of feeder cells and ROCK inhibitor,but undergo differentiation once injected into SCID mice. Genomic analyses,including SNP and INDEL,identified genes mutated in tumor cells,including components of apoptosis,cell attachment,and hypoxia pathways. The use of matched patient-derived cells provides a unique in vitro model for studies of early prostate cancer. View Publication

RAG2 severe combined immune deficiency (RAG2-SCID) is a lethal disorder caused by the absence of functional T and B cells due to a differentiation block. Here,we generated induced pluripotent stem cells (iPSCs) from a RAG2-SCID patient to study the nature of the T cell developmental blockade. We observed a strongly reduced capacity to differentiate at every investigated stage of T cell development,from early CD7-CD5- to CD4+CD8+. The impaired differentiation was accompanied by an increase in CD7-CD56+CD33+ natural killer (NK) cell-like cells. T cell receptor D rearrangements were completely absent in RAG2SCID cells,whereas the rare T cell receptor B rearrangements were likely the result of illegitimate rearrangements. Repair of RAG2 restored the capacity to induce T cell receptor rearrangements,normalized T cell development,and corrected the NK cell-like phenotype. In conclusion,we succeeded in generating an iPSC-based RAG2-SCID model,which enabled the identification of previously unrecognized disorder-related T cell developmental roadblocks. View Publication