参考文献

Melanoma brain metastasis is linked to dismal prognosis and low overall survival and is detected in up to 80% of patients at autopsy. Circulating tumor cells (CTC) are the smallest functional units of cancer and precursors of fatal metastasis. We previously used an unbiased multilevel approach to discover a unique ribosomal protein large/small subunit (RPL/RPS) CTC gene signature associated with melanoma brain metastasis. In this study,we hypothesized that CTC-driven melanoma brain metastasis secondary metastasis (“metastasis of metastasis” per clinical scenarios) has targeted organ specificity for the liver. We injected parallel cohorts of immunodeficient and newly developed humanized NBSGW (huNBSGW) mice with cells from CTC-derived melanoma brain metastasis to identify secondary metastatic patterns. We found the presence of a melanoma brain–liver metastasis axis in huNBSGW mice. Furthermore,RNA sequencing analysis of tissues showed a significant upregulation of the RPL/RPS CTC gene signature linked to metastatic spread to the liver. Additional RNA sequencing of CTCs from huNBSGW blood revealed extensive CTC clustering with human B cells in these mice. CTC:B-cell clusters were also upregulated in the blood of patients with primary melanoma and maintained either in CTC-driven melanoma brain metastasis or melanoma brain metastasis CTC–derived cells promoting liver metastasis. CTC-generated tumor tissues were interrogated at single-cell gene and protein expression levels (10x Genomics Xenium and HALO spatial biology platforms,respectively). Collectively,our findings suggest that heterotypic CTC:B-cell interactions can be critical at multiple stages of metastasis. This study provides important insights into the relevance of prometastatic CTC:B-cell clusters in melanoma progression,extends the importance of the CTC RPL/RPS gene signature beyond primary metastasis/melanoma brain metastasis driving targeted organ specificity for liver metastasis (“metastasis of metastasis”),and identifies new targets for clinical melanoma metastasis therapies. View Publication

PRDX2 is significantly expressed in various cancers and is associated with the proliferation of tumor cells. Nonetheless,the precise mechanism of PRDX2 in tumor immunity remains incompletely understood. This study aims to investigate the impact of PRDX2,which is highly expressed in lung adenocarcinoma,on T cells in the tumor immune microenvironment,and its immune action target to promote the immune escape of lung cancer cells,to provide a theoretical basis for lung adenocarcinoma treatment with PRDX2 as the target. Mouse animal models to verify the effect of Conoidin A treatment on tumor growth and T cell infiltration. Flow cytometry and Western blot verified tumor cell apoptosis in the in vitro co-culture system as well as granzyme B and perforin expression in T cells. RNA-Seq was used to obtain the downstream immune molecule. si-RNA knockdown of Galectin-9 was co-cultured with T cells in vitro. Immunofluorescence and Western blot verified that PRDX2 regulates Galectin-9 expression through HDAC3. PRDX2 expression was negatively correlated with CD8 + T cell expression in LUAD patients. Inhibition of PRDX2 significantly enhanced T-cell killing of LUAD cells and reduced tumor load in both in vitro and in vivo models. Mechanistically,Conoidin A or shRNA_PRDX2 decreased Galectin-9 expression by down-regulating the phosphorylation of HDAC3,consequently enhancing the infiltration and function of CD8 + T cells. This study reveals the role of the PRDX2/HDAC3/Galectin-9 axis in LUAD immune escape and indicates Galectin-9 as a promising target for immunotherapy. The online version contains supplementary material available at 10.1186/s12967-024-05888-z. View Publication

Pseudomonas aeruginosa is a Gram-negative bacterium that is notorious for airway infections in cystic fibrosis (CF) subjects. Bacterial quorum sensing (QS) coordinates virulence factor expression and biofilm formation at population level. Better understanding of QS in the bacterium-host interaction is required. Here,we set up a new P. aeruginosa infection model,using 2D upper airway nasal organoids that were derived from 3D organoids. Using dual RNA-sequencing,we dissected the interaction between organoid epithelial cells and WT or QS-mutant P. aeruginosa strains. Since only a single healthy individual and a single CF subject were used as donors for the organoids,conclusions about CF-specific effects could not be deduced. However,P. aeruginosa induced epithelial inflammation,whereas QS signaling did not affect the epithelial airway cells. Conversely,the epithelium influenced infection-related processes of P. aeruginosa,including QS-mediated regulation. Comparison of our model with samples from the airways of CF subjects indicated that our model recapitulates important aspects of infection in vivo. Hence,the 2D airway organoid infection model is relevant and may help to reduce the future burden of P. aeruginosa infections in CF. The online version contains supplementary material available at 10.1038/s41598-024-82500-w. View Publication

Many respiratory viruses attack the airway epithelium and cause a wide spectrum of diseases for which we have limited therapies. To date,a few primary human stem cell-based models of the proximal airway have been reported for drug discovery but scaling them up to a higher throughput platform remains a significant challenge. As a result,most of the drug screening assays for respiratory viruses are performed on commercial cell line-based 2D cultures that provide limited translational ability. We optimized a primary human stem cell-based mucociliary airway epithelium model of SARS-CoV-2 infection,in 96-well air–liquid-interface (ALI) format,which is amenable to moderate throughput drug screening. We tested the model against SARS-CoV-2 parental strain (Wuhan) and variants Beta,Delta,and Omicron. We applied this model to screen 2100 compounds from targeted drug libraries using a high throughput-high content image-based quantification method. The model recapitulated the heterogeneity of infection among patients with SARS-CoV-2 parental strain and variants. While there were heterogeneous responses across variants for host factor targeting compounds,the two direct-acting antivirals we tested,Remdesivir and Paxlovid,showed consistent efficacy in reducing infection across all variants and donors. Using the model,we characterized a new antiviral drug effective against both the parental strain and the Omicron variant. This study demonstrates that the 96-well ALI model of primary human mucociliary epithelium can recapitulate the heterogeneity of infection among different donors and SARS-CoV-2 variants and can be used for moderate throughput screening. Compounds that target host factors showed variability among patients in response to SARS-CoV-2,while direct-acting antivirals were effective against SARS-CoV-2 despite the heterogeneity of patients tested. View Publication

LRRK2 is a kinase whose activity is linked to Parkinson’s disease. This study identifies a pathway that links LRRK2 activation to lysosome perturbations. This pathway involves the process known as CASM and culminates in an interaction between LRRK2 and GABARAP at the surface of lysosomes. View Publication

Borrelia (or Borreliella ) burgdorferi,the causative agent of Lyme disease,is a motile and invasive zoonotic pathogen adept at navigating between its arthropod vector and mammalian host. While motility and chemotaxis are well known to be essential for its enzootic cycle,the role of each methyl-accepting chemotaxis proteins (MCPs) in the infectious cycle of B . burgdorferi remains unclear. In this study,we show that mcp5,a gene encoding one of the most abundant MCPs in B . burgdorferi,is differentially expressed in response to environmental signals and at distinct stages of the pathogen’s enzootic cycle. Notably,mcp5 expression is regulated by the Hk1-Rrp1 and Rrp2-RpoN-RpoS pathways,two key regulatory pathways that are critical for the spirochete’s colonization of the tick vector and mammalian host,respectively. Infection experiments with an mcp5 mutant revealed that spirochetes lacking MCP5 were unable to establish infections in either C3H/HeN mice or Severe Combined Immunodeficiency (SCID) mice,which are deficient in adaptive immunity,underscoring MCP5’s critical role in mammalian infection. However,the mcp5 mutant was able to establish infection and disseminate in NOD SCID Gamma (NSG) mice,which are deficient in both adaptive and most innate immune responses,suggesting that MCP5 plays an important role in evading host innate immunity. Moreover,NK cell depletion in C3H and SCID mice restored the infectivity of the mcp5 mutant,further highlighting MCP5’s role in evading NK cell-associated immunity. Co-culture assays with NK cells and macrophages revealed that the mcp5 mutant enhanced interferon-gamma production by NK cells. In the tick vector,the mcp5 mutants survived feeding but failed to transmit to mice. These findings reveal that MCP5,regulated by both the Rrp1 and Rrp2 pathways,is critical for establishing infection in mammalian hosts by evading NK cell-mediated host innate immunity and is important for the transmission of spirochetes from ticks to mammalian hosts. This work provides a foundation for further elucidation of chemotactic signals sensed by MCP5 that facilitate B . burgdorferi in evading host defenses. View Publication

The only cure of HIV has been achieved in a small number of people who received a hematopoietic stem cell transplant (HSCT) comprising allogeneic cells carrying a rare,naturally occurring,homozygous deletion in the CCR5 gene. The rarity of the mutation and the significant morbidity and mortality of such allogeneic transplants precludes widespread adoption of this HIV cure. Here,we show the application of CRISPR/Cas9 to achieve >90% CCR5 editing in human,mobilized hematopoietic stem progenitor cells (HSPC),resulting in a transplant that undergoes normal hematopoiesis,produces CCR5 null T cells,and renders xenograft mice refractory to HIV infection. Titration studies transplanting decreasing frequencies of CCR5 edited HSPCs demonstrate that <90% CCR5 editing confers decreasing protective benefit that becomes negligible between 54% and 26%. Our study demonstrates the feasibility of using CRISPR/Cas9/RNP to produce an HSPC transplant with high frequency CCR5 editing that is refractory to HIV replication. These results raise the potential of using CRISPR/Cas9 to produce a curative autologous HSCT and bring us closer to the development of a cure for HIV infection. Subject terms: HIV infections,CRISPR-Cas9 genome editing,Retrovirus,Translational research View Publication

Introduction: Pathogenic variants in the OFD1 gene are linked to a spectrum of syndromes that exhibit partial clinical overlap. Hemizygous loss-of-function variants are considered lethal in males,while heterozygous loss-of-function variants generally result in oro-facial-digital syndrome type 1. A reported phenotype,Simpson–Golabi–Behmel syndrome type 2,was published once but remains controversial,with many specialists questioning its validity and arguing about its continued listing in the OMIM database. Methods: To investigate the genetic and phenotypic characteristics of the patients,we performed clinical exome sequencing,family-based genetic analysis,X-inactivation studies,electron microscopy,and detailed clinical assessments. Results: Three patients from unrelated families carrying loss-of-function variants in the OFD1 gene were identified,emphasizing the diverse phenotypic spectrum of OFD1 -associated disorders. The first patient,a female with a heterozygous frameshift variant p.(Gln398LeufsTer2),was diagnosed with oro-facial-digital syndrome type 1. The second patient,a male with a heterozygous nonsense variant p.(Gln892Ter),presented with features resembling Simpson–Golabi–Behmel syndrome type 2,as previously reported under this diagnosis. The third patient,a male with another heterozygous nonsense variant p.(Glu879Ter),exhibited isolated primary ciliary dyskinesia without any syndromic features. Conclusions: This study contributes to the growing body of evidence on the expanding phenotypic spectrum of OFD1 -associated disorders. It underscores the need for further investigation into the molecular mechanisms underlying the diverse presentations and the necessity of re-evaluating diagnostic classifications for conditions such as SGBS2 in the context of variants in the OFD1 gene. View Publication

Cancer stem cells (CSCs) play a key role in non-small cell lung cancer (NSCLC) chemoresistance and metastasis. In this study,we used two NSCLC cell lines to investigate the regulating effect of hypoxia in the induction and maintenance of CSC traits. Our study demonstrated hypoxia-induced stemness and chemoresistance at levels comparable to those in typical CSC sphere culture. Activation of the NF-κB pathway (by transfection of NF-κB-p65) plays a key role in NSCLC CSCs and chemoresistance. Disulfiram (DS),an anti-alcoholism drug,showed a strong in vitro anti-CSC effect. It blocked cancer cell sphere reformation and clonogenicity,synergistically enhanced the cytotoxicity of four anti-NSCLC drugs (doxorubicin,gemcitabine,oxaliplatin and paclitaxel) and reversed hypoxia-induced resistance. The effect of DS on CSCs is copper-dependent. A very short half-life in the bloodstream is the major limitation for the translation of DS into a cancer treatment. Our team previously developed a poly lactic-co-glycolic acid (PLGA) nanoparticle encapsulated DS (DS-PLGA) with a long half-life in the bloodstream. Intra venous injection of DS-PLGA in combination with the oral application of copper gluconate has strong anticancer efficacy in a metastatic NSCLC mouse model. Further study may be able to translate DS-PLGA into cancer applications. View Publication

Previous studies have indicated that the presence of cancer stem cells may be a contributing factor to the development of metastasis in colorectal cancer patients. Cancer stem cells represent a small subpopulation within the tumor mass that exhibits heightened resistance to treatment and possesses the capacity for self-replication,epithelial–mesenchymal transition,and the generation of new tumors. The tumor microenvironment secretes and releases several molecules that facilitate the self-renewal of cancer stem cells and provide support for colorectal cancer progression. microRNAs are involved in direct cell-to-cell signaling and paracrine signaling between tumor cells and other tumor microenvironment components. They could act as tumor suppressors or oncomiRs,and their deregulation is involved in colorectal cancer progression and cancer stem cell formation. In our previous studies,we demonstrated the oncosuppressive function of miR-486-5p in colorectal cancer; these findings prompted us to conduct a more detailed investigation into its role in cancer stem cell phenotypes. Background: Colorectal cancer (CRC) is the third diagnosed cancer worldwide. Forty-four percent of metastatic colorectal cancer patients were diagnosed at an early stage. Despite curative resection,approximately 40% of patients will develop metastases within a few years. Previous studies indicate the presence of cancer stem cells (CSCs) and their contribution to CRC progression and metastasis. miRNAs deregulation plays a role in CSCs formation and in tumor development. In light of previous studies,we investigated the role of miR-486-5p to understand its role in CSC better. Methods: The expression of miR-486-5p was assessed in adherent cells and spheres generated from two CRC cell lines to observe the difference in expression in CSC-enriched spheroids. Afterward,we overexpressed and underexpressed this miRNA in adherent and sphere cultures through the transfection of a miR-486-5p mimic and a mimic inhibitor. Results: The results demonstrated that miR-486-5p exhibited a notable downregulation in CSC models,and its overexpression led to a significant decrease in colony size. Conclusions: In this study,we confirmed that miR-486-5p plays an oncosuppressive role in CRC,thereby advancing our understanding of the role of this microRNA in the CSC phenotype. View Publication