ObjectiveIntrahepatic cholangiocarcinoma (ICC)—a rare liver malignancy with limited therapeutic o... more ObjectiveIntrahepatic cholangiocarcinoma (ICC)—a rare liver malignancy with limited therapeutic options—is characterised by aggressive progression, desmoplasia and vascular abnormalities. The aim of this study was to determine the role of placental growth factor (PlGF) in ICC progression.DesignWe evaluated the expression of PlGF in specimens from ICC patients and assessed the therapeutic effect of genetic or pharmacologic inhibition of PlGF in orthotopically grafted ICC mouse models. We evaluated the impact of PlGF stimulation or blockade in ICC cells and cancer-associated fibroblasts (CAFs) using in vitro 3-D coculture systems.ResultsPlGF levels were elevated in human ICC stromal cells and circulating blood plasma and were associated with disease progression. Single-cell RNA sequencing showed that the major impact of PlGF blockade in mice was enrichment of quiescent CAFs, characterised by high gene transcription levels related to the Akt pathway, glycolysis and hypoxia signalling. ...
Proceedings of the National Academy of Sciences, 2020
Significance The burgeoning intestinal microbiota pose a unique challenge to the developing immun... more Significance The burgeoning intestinal microbiota pose a unique challenge to the developing immune system of the newborn. Extensive crosstalk between mucosal immune cells and microbes is necessary to establish healthy microbial communities and promote the development of productive mucosal immunity. However, it is yet unknown whether intestinal microbes also influence the development of lymphocytes themselves in the thymus. Here, we report that thymic distribution of transcription factor PLZF-expressing innate lymphocytes is regulated by intestinal microbes in early life. Migratory plasmacytoid dendritic cells are one conveyor of microbial information to developing thymic cells and perturbation of this entero-thymic communication in early life impacts disease susceptibility in adulthood. Our data highlight a novel regulation of thymic lymphocyte development by early-life microbiota.
Osteosarcoma is the most common bone cancer in children and adolescents. Previously, we have foun... more Osteosarcoma is the most common bone cancer in children and adolescents. Previously, we have found that cyclin-dependent kinase 11 (CDK11) signaling was essential for osteosarcoma cell growth and survival. Subsequently, CDK11 siRNA gene targeting, expression profiling, and network reconstruction of differentially expressed genes were performed between CDK11 knock down and wild type osteosarcoma cells. Reconstructed network of the differentially expressed genes pointed to the AR as key to CDK11 signaling in osteosarcoma. CDK11 increased transcriptional activation of AR gene in osteosarcoma cell lines. AR protein was highly expressed in various osteosarcoma cell lines and patient tumor tissues. Tissue microarray analysis showed that the disease-free survival rate for patients with high-expression of AR was significantly shorter than for patients with low-expression of AR. In addition, AR gene expression knockdown via siRNA greatly inhibited cell growth and viability. Similar results w...
Proteins bearing short sequence tags (ExoTags) can be specifically modified in a reaction catalyz... more Proteins bearing short sequence tags (ExoTags) can be specifically modified in a reaction catalyzed by the ADP-ribosyltransferase of Pseudomonas ExoT. ExoT tolerates a wide range of substituents at the adenosine N6 position, allowing the convenient installation of chemical diversity at the site of the tag. The ExoTag can be placed at either the N - or the C -terminus of the protein of interest or internally. The minimum tag is Arg-Leu-Ser-Arg (RLSR). Tandem copies of the minimal ExoTag enhance the efficiency of modification and allow modification at multiple sites within the tag. ExoT can be used to conjugate NAD-RNA to ExoTag bearing proteins, providing a convenient method for introducing nucleic acid bar codes for highly sensitive detection of proteins by polymerase chain reaction.
ABSTRACTOsteosarcoma is the most common type primary malignant tumor of bone. Patients with regio... more ABSTRACTOsteosarcoma is the most common type primary malignant tumor of bone. Patients with regional osteosarcoma are routinely treated with surgery and chemotherapy. In addition, many patients with metastatic or recurrent osteosarcoma show poor prognosis with current chemotherapy agents. Therefore, it is important to improve the general condition and the overall survival rate of patients with osteosarcoma by identifying novel therapeutic strategies. Recent studies have revealed that CDK11 is essential in osteosarcoma cell growth and survival by inhibiting CDK11 mRNA expression with RNAi. Here, we apply the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)‐Cas9 system, a robust and highly efficient novel genome editing tool, to determine the effect of targeting endogenous CDK11 gene at the DNA level in osteosarcoma cell lines. We show that CDK11 can be efficiently silenced by CRISPR‐Cas9. Inhibition of CDK11 is associated with decreased cell proliferation and viabil...
A correction to this article has been published and is linked from the HTML and PDF versions of t... more A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has not been fixed in the paper.
Proceedings of the National Academy of Sciences of the United States of America, Jan 22, 2015
Analysis of embryonic fibroblasts from GFP reporter mice indicates that the fibroblast cell type ... more Analysis of embryonic fibroblasts from GFP reporter mice indicates that the fibroblast cell type harbors a large collection of developmentally and phenotypically heterogeneous subtypes. Some of these cells exhibit multipotency, whereas others do not. Multiparameter flow cytometry analysis shows that a large number of distinct populations of fibroblast-like cells can be found in cultures initiated from different embryonic organs, and cells sorted according to their surface phenotype typically retain their characteristics on continued propagation in culture. Similarly, surface phenotypes of individual cloned fibroblast-like cells exhibit significant variation. The fibroblast cell class appears to contain a very large number of denumerable subtypes.
ObjectiveIntrahepatic cholangiocarcinoma (ICC)—a rare liver malignancy with limited therapeutic o... more ObjectiveIntrahepatic cholangiocarcinoma (ICC)—a rare liver malignancy with limited therapeutic options—is characterised by aggressive progression, desmoplasia and vascular abnormalities. The aim of this study was to determine the role of placental growth factor (PlGF) in ICC progression.DesignWe evaluated the expression of PlGF in specimens from ICC patients and assessed the therapeutic effect of genetic or pharmacologic inhibition of PlGF in orthotopically grafted ICC mouse models. We evaluated the impact of PlGF stimulation or blockade in ICC cells and cancer-associated fibroblasts (CAFs) using in vitro 3-D coculture systems.ResultsPlGF levels were elevated in human ICC stromal cells and circulating blood plasma and were associated with disease progression. Single-cell RNA sequencing showed that the major impact of PlGF blockade in mice was enrichment of quiescent CAFs, characterised by high gene transcription levels related to the Akt pathway, glycolysis and hypoxia signalling. ...
Proceedings of the National Academy of Sciences, 2020
Significance The burgeoning intestinal microbiota pose a unique challenge to the developing immun... more Significance The burgeoning intestinal microbiota pose a unique challenge to the developing immune system of the newborn. Extensive crosstalk between mucosal immune cells and microbes is necessary to establish healthy microbial communities and promote the development of productive mucosal immunity. However, it is yet unknown whether intestinal microbes also influence the development of lymphocytes themselves in the thymus. Here, we report that thymic distribution of transcription factor PLZF-expressing innate lymphocytes is regulated by intestinal microbes in early life. Migratory plasmacytoid dendritic cells are one conveyor of microbial information to developing thymic cells and perturbation of this entero-thymic communication in early life impacts disease susceptibility in adulthood. Our data highlight a novel regulation of thymic lymphocyte development by early-life microbiota.
Osteosarcoma is the most common bone cancer in children and adolescents. Previously, we have foun... more Osteosarcoma is the most common bone cancer in children and adolescents. Previously, we have found that cyclin-dependent kinase 11 (CDK11) signaling was essential for osteosarcoma cell growth and survival. Subsequently, CDK11 siRNA gene targeting, expression profiling, and network reconstruction of differentially expressed genes were performed between CDK11 knock down and wild type osteosarcoma cells. Reconstructed network of the differentially expressed genes pointed to the AR as key to CDK11 signaling in osteosarcoma. CDK11 increased transcriptional activation of AR gene in osteosarcoma cell lines. AR protein was highly expressed in various osteosarcoma cell lines and patient tumor tissues. Tissue microarray analysis showed that the disease-free survival rate for patients with high-expression of AR was significantly shorter than for patients with low-expression of AR. In addition, AR gene expression knockdown via siRNA greatly inhibited cell growth and viability. Similar results w...
Proteins bearing short sequence tags (ExoTags) can be specifically modified in a reaction catalyz... more Proteins bearing short sequence tags (ExoTags) can be specifically modified in a reaction catalyzed by the ADP-ribosyltransferase of Pseudomonas ExoT. ExoT tolerates a wide range of substituents at the adenosine N6 position, allowing the convenient installation of chemical diversity at the site of the tag. The ExoTag can be placed at either the N - or the C -terminus of the protein of interest or internally. The minimum tag is Arg-Leu-Ser-Arg (RLSR). Tandem copies of the minimal ExoTag enhance the efficiency of modification and allow modification at multiple sites within the tag. ExoT can be used to conjugate NAD-RNA to ExoTag bearing proteins, providing a convenient method for introducing nucleic acid bar codes for highly sensitive detection of proteins by polymerase chain reaction.
ABSTRACTOsteosarcoma is the most common type primary malignant tumor of bone. Patients with regio... more ABSTRACTOsteosarcoma is the most common type primary malignant tumor of bone. Patients with regional osteosarcoma are routinely treated with surgery and chemotherapy. In addition, many patients with metastatic or recurrent osteosarcoma show poor prognosis with current chemotherapy agents. Therefore, it is important to improve the general condition and the overall survival rate of patients with osteosarcoma by identifying novel therapeutic strategies. Recent studies have revealed that CDK11 is essential in osteosarcoma cell growth and survival by inhibiting CDK11 mRNA expression with RNAi. Here, we apply the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)‐Cas9 system, a robust and highly efficient novel genome editing tool, to determine the effect of targeting endogenous CDK11 gene at the DNA level in osteosarcoma cell lines. We show that CDK11 can be efficiently silenced by CRISPR‐Cas9. Inhibition of CDK11 is associated with decreased cell proliferation and viabil...
A correction to this article has been published and is linked from the HTML and PDF versions of t... more A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has not been fixed in the paper.
Proceedings of the National Academy of Sciences of the United States of America, Jan 22, 2015
Analysis of embryonic fibroblasts from GFP reporter mice indicates that the fibroblast cell type ... more Analysis of embryonic fibroblasts from GFP reporter mice indicates that the fibroblast cell type harbors a large collection of developmentally and phenotypically heterogeneous subtypes. Some of these cells exhibit multipotency, whereas others do not. Multiparameter flow cytometry analysis shows that a large number of distinct populations of fibroblast-like cells can be found in cultures initiated from different embryonic organs, and cells sorted according to their surface phenotype typically retain their characteristics on continued propagation in culture. Similarly, surface phenotypes of individual cloned fibroblast-like cells exhibit significant variation. The fibroblast cell class appears to contain a very large number of denumerable subtypes.
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