rb1 tm2brn j  (Jackson Laboratory)

Journal: Cancer Discovery

Article Title: Serotonin Modulates Lineage Plasticity in Neuroendocrine Prostate Cancer via Epigenetic Reprogramming

doi: 10.1158/2159-8290.CD-25-0974

Figure Lengend Snippet: Identification of serotonin signaling as a candidate target in NEPC through high-throughput compound screening and clinical transcriptomic data. A, A schematic showing the compound screening pipeline using the NEPC cell line LASCPC-01 and the CellTiter-Glo viability assay. B, A waterfall plot showing the relative viability of NEPC cells upon treatment with 1,112 FDA-approved compounds. C, Mechanism of action enrichment analysis from the Drug Repurposing Hub highlighting serotonin reuptake inhibitors as the top enriched class among active compounds. D, Serotonin-related targets identified from Drug Central among the top enriched hits. E, Uniform Manifold Approximation and Projection for Dimension Reduction (UMAP) plot of scRNA-seq data from CRPC epithelial cells , showing NE cell clusters and SOX2 expression. F, A Venn diagram showing NE marker genes shared between human CRPC single-cell data and the TKO (Pb-Cre4: Pten f/f ; Trp53 f/f ; Rb1 f/f ) NEPC mouse model . Ranking of shared NE genes in the original Beltran and colleagues dataset . G, UMAP subclustering of epithelial cells showing SOX2 + DDC + , SOX2 + DDC − , and SOX2 − DDC − populations. H, A gene expression correlation heatmap between NE markers, AR pathway genes, and DDC / SLC6A4 using bulk RNA-seq data (Beltran and colleagues cohort; ref. ). I, Box plots showing expression levels of DDC and SLC6A4 in four public prostate cancer datasets (Beltran and colleagues, Tzelepi and colleagues, SU2C/Abida and colleagues, and Taylor and colleagues cohorts; refs. – ). J, Representative hematoxylin and eosin (H&E) and immunohistochemistry (IHC) staining of DDC, AR, and CHGA in prostate tissues from benign prostate hyperplasia (BPH, n = 10), hormone-sensitive prostate cancer (HSPC, n = 27), CRPC ( n = 13), and NEPC ( n = 15) patients. Scale bar, 100 μm. K, Representative H&E and IHC staining of DDC, AR, and CHGA in prostate, liver, and lung tumors from TKO and DKO (Pb-Cre4: Pten f/f ; Trp53 f/f ) mice. Scale bar, 100 μm.

Article Snippet: Tg (Pbsn-cre)4Prb/J (026662, RRID: IMSR_JAX:026662), B6.129P2- Trp53 tm1Brn /J (008462, RRID: IMSR_JAX:008462), Rb1 tm2Brn /J (026563, RRID: IMSR_JAX:026563), and B6.129S4- Pten tm1Hwu /J mice (006440, RRID: IMSR_JAX:006440) were bought from The Jackson Laboratory to generate TKO mice (Pb-Cre4: Pten f/f ; Trp53 f/f ; Rb1 f/f ), DKO mice (Pb-Cre4: Pten f/f ; Trp53 f/f ), and Pten f/f ; Trp53 f/f ; Rb1 f/f mice.

Techniques: High Throughput Screening Assay, Viability Assay, Expressing, Marker, Single Cell, Gene Expression, RNA Sequencing, Immunohistochemistry

Journal: Nature Communications

Article Title: The ELAVL3/MYCN positive feedback loop provides a therapeutic target for neuroendocrine prostate cancer

doi: 10.1038/s41467-023-43676-3

Figure Lengend Snippet: a Venn diagram showing the overlap of upregulated differentially expressed genes for the four NEPC RNA-seq datasets: (1) neuroendocrine prostate cancer (NEPC) versus castration-resistant prostate cancer (CRPC), data obtained from Beltran 2016 cohort ; (2) Pb-Cre4: Pten f/f ; LSL- MYCN +/+ versus Pb-Cre4: Pten f/f ; LSL- MYCN −/− organoids, data obtained from GSE86532 ; (3) Pb-Cre4: Pten f/f ; Trp53 f/f ; Rb1 f/f versus Pb-Cre4: Pten f/f , data obtained from GSE90891 ; (4) two NE-high single-cell clusters versus 14 NE-low single-cell clusters, data obtained from GSE137829 . Common genes across all analytes are indicated. b Dot plots illustrating the Pearson correlation analysis between the expression of ELAVL3 , NE-related genes, and AR-related genes. Data obtained from Beltran 2016 cohort . c Representative H&E and immunofluorescence staining of AR, SYP, CHGA, and ELAVL3 in prostate tissues from indicated patient groups (BPH ( n = 15), HSPC ( n = 144), CRPC ( n = 15), and NEPC ( n = 22)). Scale bar, 50 μm. d Quantification of ELAVL3 staining in prostate tissues from indicated patient groups, each dot represents tissue from an individual patient. e QPCR showing relative mRNA expression of ELAVL3 , NE-related genes ( MYCN , SOX2 , CHGA , CHGB , NCAM1 , SYP , and NSE ), and AR in a panel of prostate cancer cells ( n = 3 biologically independent experiments). f Western blot showing protein expression of ELAVL3 and NE-related genes (MYCN, NSE, SYP, and NCAM1) in a panel of prostate cancer cells. g Representative H&E and immunofluorescence staining of Elavl3, Syp, Ncam1, Ar, and Ki67 in tumors from mice of Pb-Cre4: Pten f/f ; Trp53 f/f ( n = 10) and Pb-Cre4: Pten f/f ; Trp53 f/f ; Rb1 f/f ( n = 10). Scale bar, 50 μm. h Quantification of Elavl3 staining in prostate tumor tissues from mice groups: Pb-Cre4: Pten f/f ; Trp53 f/f ( n = 10) and Pb-Cre4: Pten f/f ; Trp53 f/f ; Rb1 f/f ( n = 10); each dot represents tissue from an individual mouse. Data presented as mean ± s.e.m ( d , e , h ). Statistical significance was determined by two-tailed unpaired Student’s t -test ( e , h ) or one-way analysis of variance (ANOVA) with Dunnett’s multiple comparisons ( d , e ). Western blot experiments were repeated three times independently, with similar results ( f ). NE-related, related to neuroendocrine differentiation. AR-related, related to the androgen receptor pathway. BPH benign prostatic hyperplasia. HSPC hormone-sensitive prostate cancer. Source data are provided as a Source Data file.

Article Snippet: Tg(Pbsn-cre)4Prb/J (026662), B6.129P2- Trp53 tm1Brn /J (008462), Rb1 tm2Brn /J (026563), and B6.129S4- Pten tm1Hwu /J mice (006440) were bought from Jackson Laboratory to generate Pb-Cre4: Pten f /f ; Trp53 f/f ; Rb1 f/f mice, Pb-Cre4: Pten f/f ; Trp53 f/f mice, and Pten f/f ; Trp53 f/f ; Rb1 f/f mice.

Techniques: RNA Sequencing, Expressing, Immunofluorescence, Staining, Western Blot, Two Tailed Test

Journal: Nature Communications

Article Title: The ELAVL3/MYCN positive feedback loop provides a therapeutic target for neuroendocrine prostate cancer

doi: 10.1038/s41467-023-43676-3

Figure Lengend Snippet: a Photograph showing morphology of organoids from Pb-Cre4: Pten f/f ; Trp53 f/f (DKO) and Pb-Cre4: Pten f/f ; Trp53 f/f ; Rb1 f/f (TKO) prostate tumors treated with PBS or PP. Scale bar, 50 μm. b Cell viability of organoids from DKO and TKO prostate tumors treated with/without PP ( n = 5 independent treated). c Schematic illustration showing the treatment and PET/CT imaging of TKO. d [ 18 F]-FDG PET/CT imaging of TKO before and after indicated treatment. e Quantification of [ 18 F]-FDG PET/CT signal by maximum standardized uptake value in TKO before and after indicated treatment ( n = 3 per group). f Kaplan–Meier survival analysis of TKO treated with PP or PBS (PBS, n = 14; PP, n = 11). g , h Representative immunofluorescence staining and quantification of EdU and Elavl3 in tumors from TKO treated with PP or PBS. Scale bar, 50 μm. (PBS, n = 9; PP, n = 8). i , j Representative H&E, immunohistochemistry staining and quantification of Ki67 and cleaved-caspase 3 in tumors from TKO treated with PP or PBS. Scale bar, 50 μm. ( n = 9 per group). k Representative H&E staining of lung and liver metastases from TKO treated with PP or PBS ( n = 3 per group). Scale bar, 2 mm. l Summary of the liver and lung metastases from TKO treated with PP or PBS. m , n QPCR showing relative mRNA expression of indicated genes and western blot showing indicated protein expression in tumor tissues from TKO treated with PP or PBS ( n = 3 per group). o Schematic showing that ELAVL3 is transcriptionally activated by MYCN, and then in turn binds to and stabilizes the mRNA of MYCN and RICTOR . ELAVL3 can be released into EVs and induce neuroendocrine differentiation of recipient cells. PP disrupts the interaction of ELAVL3 and MYCN, reduces ELAVL3-induced neuroendocrine differentiation, and provides a promising drug repurposing strategy for the treatment of NEPC. Data presented as mean ± s.e.m. ( b , e , h , j , m ). Statistical significance was determined by two-tailed unpaired Student’s t -test ( b , e , h , j , m ), or log-rank (Mantel-Cox) test ( f ). Western blot experiments were repeated three times independently, with similar results ( n ). Source data are provided as a Source Data file.

Article Snippet: Tg(Pbsn-cre)4Prb/J (026662), B6.129P2- Trp53 tm1Brn /J (008462), Rb1 tm2Brn /J (026563), and B6.129S4- Pten tm1Hwu /J mice (006440) were bought from Jackson Laboratory to generate Pb-Cre4: Pten f /f ; Trp53 f/f ; Rb1 f/f mice, Pb-Cre4: Pten f/f ; Trp53 f/f mice, and Pten f/f ; Trp53 f/f ; Rb1 f/f mice.

Techniques: Positron Emission Tomography-Computed Tomography, Imaging, Immunofluorescence, Staining, Immunohistochemistry, Expressing, Western Blot, Two Tailed Test

Journal: Cell Death and Differentiation

Article Title: Disruption of GMNC-MCIDAS multiciliogenesis program is critical in choroid plexus carcinoma development

doi: 10.1038/s41418-022-00950-z

Figure Lengend Snippet: A Combined activation of NOTCH and SHH signaling, or loss of Rb1 / Trp53 tumor suppressors drives CPC development. B GMNC-MCIDAS program mediates multiciliation in CP epithelium, and is repressed by NOTCH signaling in roof plate progenitors, whereas NOTCH inhibitor IMR-1 promotes GMNC-dependent multiciliation and suppresses tumor growth. GMNC-MCIDAS program suppression in Trp53 -deficient CPC maintains monociliated tumor cells.

Article Snippet: Gt(ROSA)26Sor tm1.Notch1Dam /J ( Rosa26-NICD1 ) mice, B6N.129- Ptch1 tm1Hahn /J ( Ptch flox/flox ) mice, B6.129P2- Trp53 tm1Brn /J ( Trp53 flox/flox ) mice, Rb1 tm2Brn /J ( Rb flox/flox ) mice, and C57BL/6 mice (all from Jackson Laboratory, Bar Harbor, ME, USA), Tg(Lmx1a-cre)1Kjmi ( Lmx1a-Cre ) mice, Gmnc tm1Strc ( Gemc1 −/+ ) mice, and Gmnc tm1.1Strc ( Gemc1 flox/+ ) mice were maintained by breeding with C57BL/6 mice (Supplementary Table ).

Techniques: Activation Assay