Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: RABL6A is an essential driver of MPNSTs that negatively regulates the RB1 pathway and sensitizes tumor cells to CDK4/6 inhibitors

doi: 10.1158/1078-0432.CCR-19-2706

Figure Lengend Snippet: (A) Immunohistochemical (IHC) staining of RABL6A in tissue microarrays (TMAs) of patient-matched NF (mainly plexiform NFs) and MPNSTs. Images at 10X magnification. Higher magnification images (400X) of RABL6A (B) and p27 (D) IHC staining of NF, ANNUBP and MPNST cores from the TMA. Stain intensity indicated in parentheses. H-scores, representing the stain intensity × percent of cells stained, of RABL6A (C) and p27 (E) in NF, ANNUBP and MPNST samples. Error bars, SEM. P-value, Student’s t-test. *, p<0.05; **, p<0.01. (F) Heat-map of RNA-Seq data with hierarchical clustering of a subset of functionally related differentially expressed genes (n=110) between paired patient NFs, ANNUBPs (designated A) and MPNSTs. Genes were categorized by IPA software. Red, relatively increased expression; blue, relatively decreased expression. Genes within each group are listed in Supplemental Table 2. (G) An MPNST gene activity wheel, depicting select genes associated with RABL6A expression, shows MPNSTs have an activated ‘RABL6A signature’ of gene expression (22, 23). Transcript levels of the indicated genes in MPNSTs: decreased (blue), increased (red), unchanged (black). Circled genes reflect pathway activation (red) or inhibition (blue). Expression of RABL6A is associated with increased (red arrows) or decreased (blue bars) mRNA levels of the displayed genes.

Article Snippet: Three well characterized human MPNST cell lines were used for these studies: the sporadic human MPNST cell line STS26T (referred to as ‘26T’; NF1 wild-type; Cellosaurus RRID:CVCL_8917), NF1-associated human MPNST cell line S462 (Cellosaurus RRID:CVCL_1Y70), and NF1-associated human MPNST cell line sNF96.2 (ATCC CRL-2884).

Techniques: Immunohistochemical staining, Immunohistochemistry, Staining, RNA Sequencing, Software, Expressing, Activity Assay, Gene Expression, Activation Assay, Inhibition

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: RABL6A is an essential driver of MPNSTs that negatively regulates the RB1 pathway and sensitizes tumor cells to CDK4/6 inhibitors

doi: 10.1158/1078-0432.CCR-19-2706

Figure Lengend Snippet: S462 MPNST cells were transduced with empty vector (EV) or p27 shRNA viruses and subsequently infected with control (CON) or RABL6A shRNAs (KD1 and KD2). (A) Quantification of the fold change in cell death after RABL6A knockdown shows that p27 inactivation rescues cell death caused by RABL6A loss. (B) Representative westerns show partial (KD1) to full (KD2) restoration of RB1 phosphorylation (p-RB1) upon co-depletion of p27 with RABL6A. Below, cell ratio (relative to control cells) for the displayed experiment. (C) ImageJ quantification of western data from three or more experiments showing partial to full restoration of phospho-RB1 when p27 is depleted in tandem with RABL6A. (D) Representative images of EdU staining show p27 inactivation rescues DNA synthesis in RABL6A depleted cells. Percent EdU positive cells (cyan colored nuclei versus dark blue nuclei for EdU negative cells) are denoted. (E) Quantification of EdU positivity from three independent experiments. Data in A, C, and E are from three of more independent experiments. Error bars, SD from mean. P-value, Student’s t-test with Bonferroni’s correction. *, p<0.05; **, p<0.01.

Article Snippet: Three well characterized human MPNST cell lines were used for these studies: the sporadic human MPNST cell line STS26T (referred to as ‘26T’; NF1 wild-type; Cellosaurus RRID:CVCL_8917), NF1-associated human MPNST cell line S462 (Cellosaurus RRID:CVCL_1Y70), and NF1-associated human MPNST cell line sNF96.2 (ATCC CRL-2884).

Techniques: Transduction, Plasmid Preparation, shRNA, Infection, Control, Knockdown, Phospho-proteomics, Western Blot, Staining, DNA Synthesis

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: RABL6A is an essential driver of MPNSTs that negatively regulates the RB1 pathway and sensitizes tumor cells to CDK4/6 inhibitors

doi: 10.1158/1078-0432.CCR-19-2706

Figure Lengend Snippet: (A) Western blots show elevated RABL6A levels in both NF1-associated (T265p21, 90–8, S520 and S462) and sporadic (STS26T [26T]c and YST-1) MPNST cell lines versus precursor, primary normal human Schwann cells (NHSC). (B) Representative westerns show that knockdown of RABL6A (KD1 and KD2) causes induction of cleaved PARP, a marker of apoptosis, in MPNST cells but not in NHSC. Loading control, GAPDH or γ-tubulin. Cell number relative to CON (cell ratio) for each cell line is shown. CON, empty vector control. (C) Quantification of cell death, as measured by trypan blue exclusion, shows RABL6A knockdown induces the death of MPNST cells but not NHSC. P-value, Student’s t-test with Dunnett’s correction for KD versus CON; **, p<0.01. ns, not significant. (D) RABL6A loss in both NHSC and MPNST cells causes reduced DNA synthesis, as measured by EdU positivity relative to CON cells (set at 1.0). **, P<0.001 for all samples compared to controls for each cell line. (E) Pie chart schematics of representative DNA content analyses show RABL6A loss causes G1 accumulation and decreased S phase. Data in B – E are from three of more independent experiments. C, D: Error bars, SD from mean.

Article Snippet: Three well characterized human MPNST cell lines were used for these studies: the sporadic human MPNST cell line STS26T (referred to as ‘26T’; NF1 wild-type; Cellosaurus RRID:CVCL_8917), NF1-associated human MPNST cell line S462 (Cellosaurus RRID:CVCL_1Y70), and NF1-associated human MPNST cell line sNF96.2 (ATCC CRL-2884).

Techniques: Western Blot, Knockdown, Marker, Control, Plasmid Preparation, DNA Synthesis

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: RABL6A is an essential driver of MPNSTs that negatively regulates the RB1 pathway and sensitizes tumor cells to CDK4/6 inhibitors

doi: 10.1158/1078-0432.CCR-19-2706

Figure Lengend Snippet: (A) Representative westerns of MPNST cell lines expressing empty vector (CON) versus RABL6A shRNAs (KD1 and KD2). RABL6A loss causes p27 upregulation and CDK2 downregulation. (B) ImageJ quantification of western data from three or more experiments showing fold increase in p27 expression following RABL6A knockdown. (C) Representative westerns show loss of RABL6A in MPNST cells causes reduced phosphorylation of RB1 at CDK4/6 targeted sites (S807/811). (D, E) ImageJ quantification of western data from three or more experiments showing reduced levels of phospho-RB1 (D) and CDK2 (E) in RABL6A knockdown cells relative to CON cells (set at 1.0). (F) Proposed model of the RABL6A-p27-RB1 pathway and how it contributes to MPNST cell proliferation and survival. Dashed arrow indicates RABL6A may also act through RB1-independent mechanisms to promote MPNST pathogenesis. Data in B, D and E are normalized to loading control or total RB1, as indicated. Error bars, SD from mean. P-value, Student’s t-test with Dunnett’s correction for each KD versus the CON (set at 1.0) for each cell line. *, p<0.05; **, p<0.01.

Article Snippet: Three well characterized human MPNST cell lines were used for these studies: the sporadic human MPNST cell line STS26T (referred to as ‘26T’; NF1 wild-type; Cellosaurus RRID:CVCL_8917), NF1-associated human MPNST cell line S462 (Cellosaurus RRID:CVCL_1Y70), and NF1-associated human MPNST cell line sNF96.2 (ATCC CRL-2884).

Techniques: Expressing, Plasmid Preparation, Western Blot, Knockdown, Phospho-proteomics, Control

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: RABL6A is an essential driver of MPNSTs that negatively regulates the RB1 pathway and sensitizes tumor cells to CDK4/6 inhibitors

doi: 10.1158/1078-0432.CCR-19-2706

Figure Lengend Snippet: (A) MPNST cells were treated for 3 days with varying concentrations of palbociclib and relative cell number assessed by CellQuant. IC50 values indicated at top right. (B) Left, MPNST cells were treated for 2 days with 10 μM palbociclib and viability assessed by trypan blue exclusion. Right, Representative westerns showing reduced p-RB1 at CDK4/6 sites after drug treatment. Below, cell ratios relative to untreated controls show palbociclib reduces MPNST cell number. (C) Relative number of primary human NHSCs after palbociclib treatment for 3 days, as measured by AlamarBlue, in low passage (≤ P4) or high passage (≥ P7) cells. IC50 value for low passage NHSC at top right. (D) Left, Percent cell death in low versus high passage NHSCs showing no induction of death by palbociclib. Right, Representative westerns verifying reduced p-RB1 S807/811 in drug treated cells. (E) S462 CON and RABL6A depleted cells (KD1 and KD2) were treated with palbociclib at varying concentrations for 3 days and relative cell number assessed by AlamarBlue. Loss of RABL6A reduces S462 cell sensitivity to palbociclib. (F) Left, Fold change in cell death reveals cells lacking RABL6A undergo reduced drug induced death compared to CON cells. Right, Representative westerns show that palbociclib effectively reduces p-RB1 807/811 in CON and KD cells. Below, ratio of p-RB1 to total RB1. Data in A-F obtained from three or more experiments. Error bars, SD from mean. B, D, F: P-value, Student’s t-test (F: with Dunnett’s correction); **, p<0.01. E: P-values determined by a generalized linear model to assess the difference between the curves.

Article Snippet: Three well characterized human MPNST cell lines were used for these studies: the sporadic human MPNST cell line STS26T (referred to as ‘26T’; NF1 wild-type; Cellosaurus RRID:CVCL_8917), NF1-associated human MPNST cell line S462 (Cellosaurus RRID:CVCL_1Y70), and NF1-associated human MPNST cell line sNF96.2 (ATCC CRL-2884).

Techniques:

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: RABL6A is an essential driver of MPNSTs that negatively regulates the RB1 pathway and sensitizes tumor cells to CDK4/6 inhibitors

doi: 10.1158/1078-0432.CCR-19-2706

Figure Lengend Snippet: (A) MPNST cells were treated for 3 days with the indicated concentrations of dinaciclib and relative cell number quantified by AlamarBlue. IC50 values are shown. (B) Contour plot of interaction index (Bliss independence model) for the combination of low doses of CDK4/6 (palbociclib) and CDK2 (dinaciclib) inhibitors in sNF96.2 cells. Red, synergy; green, antagonism. (C) Colony formation assays in sNF96.2 cells show synergism between low concentrations of dinaciclib (2 or 3 nM) and palbociclib. Left, representative images. Right, quantification of percentage area covered by cells from 3 or more experiments. P<0.0001 for all comparisons except Dina vs Palbo-125. (D) Fold change in tumor volume for sNF96.2 orthotopic tumors treated daily with vehicle (water), palbociclib (0.5 mg/dose), dinaciclib (0.16 mg/dose) or the combination. P<0.05 for all comparisons except Dina vs Combo and Dina vs Palbo, which were non-significant. (E) IHC staining for Ki-67, S100+, and phospho-RB1 S807/811 in sNF96.2 tumor samples after treatment. H&E images at 400X magnification. IHC images at 200X magnification. Quantification of (F) Ki-67, (G) phospho-RB1 S807/811, and (H) CC3 in treated tumor samples. G: *, p<0.05 for Combo vs Veh and Combo vs Dina, other comparisons not significant. H: no comparisons significant. A, C: Error bars, SD from the mean. D, F, G, H: Error bars, SEM. C, F, G, H: P-value, Student’s t-test with Bonferroni correction. D: P-values determined by a generalized linear model to assess the difference between the curves.

Article Snippet: Three well characterized human MPNST cell lines were used for these studies: the sporadic human MPNST cell line STS26T (referred to as ‘26T’; NF1 wild-type; Cellosaurus RRID:CVCL_8917), NF1-associated human MPNST cell line S462 (Cellosaurus RRID:CVCL_1Y70), and NF1-associated human MPNST cell line sNF96.2 (ATCC CRL-2884).

Techniques: Immunohistochemistry

Journal: Molecular Cancer Therapeutics

Article Title: Dual targeting of AKT and mammalian target of rapamycin: A potential therapeutic approach for malignant peripheral nerve sheath tumor

doi: 10.1158/1535-7163.mct-08-1008

Figure Lengend Snippet: Figure 1. MPNST exhibit dereg- ulation of the mTOR and AKT path- ways. A, expression of activated mTOR downstream effectors 4EBP1 (top) and S6RP (bottom) in MPNST was assessed via immuno- histochemistry. A tissue microarray consisting of human NF1-associated plexiform neurofibromas and NF1 associated as well as sporadic MPNST was used for immuno- histochemistry. A significantly higher proportion of MPNST cells expressed p4EBP1 and pS6RP compared with neurofibromas (rep- resentative tissue microarray spots are shown; original pictures were captured at ×200). Similarly, stain- ing intensity for both markers was significantly higher in the malignant tumors compared with the benign lesions. Results of Wilcoxon rank- sum tests: blue line, median of the data; bars, 25% and 75% quar- tiles; dots, mean outliers. The two lines are whisker lines, depicting the largest or smallest values that are not outliers. Statistical P values are shown. B, pAKT was evaluated as above; a significantly higher pro- portion of MPNST cells were found to exhibit a stronger pAKT expres- sion level. Furthermore, metastatic lesions expressed higher levels of pAKT compared with localized (pri- mary and recurrent) tumors.

Article Snippet: Cell Lines and Reagents Human NF1 MPNST cell lines SNF02.2 and SNF96.2 (American Type Culture Collection), ST88-14 (a gift from Dr. Jonathan Fletcher, Brigham and Women's Hospital), and T265 (a gift from Dr. George De Vries, Hines VA Hospital) and the non-NF1 sporadic human MPNST cell line STS26T (a gift from Dr. Steven Porcelli, Albert Einstein College of Medicine) were used for our studies and maintained and propagated as described previously (22).

Techniques: Expressing, Immunohistochemistry, Microarray, Staining, Whisker Assay

Journal: Molecular Cancer Therapeutics

Article Title: Dual targeting of AKT and mammalian target of rapamycin: A potential therapeutic approach for malignant peripheral nerve sheath tumor

doi: 10.1158/1535-7163.mct-08-1008

Figure Lengend Snippet: Figure 2. Human MPNSTcells exhibit activated mTOR and AKT pathways. A, Western blots showing expression of activated mTOR and its downstream targets p4EBP1 and pS6K1 in a panel of NF1-associated (SNF02.2, SNF96.2, T265, and ST88) and sporadic (STS26T) human MPNST cell lines. B, ac- tivation of AKT was also found in these cells. pAKT and its downstream substrates pGSK3β and pMDM2 are expressed in MPNST cells. C, pAKT is ex- pressed in MPNST cells when cultured in serum-free medium. D, Western blot showing expression of several possible upstream AKT modifiers in MPNST cells. Nf1, neurofibromin.

Article Snippet: Cell Lines and Reagents Human NF1 MPNST cell lines SNF02.2 and SNF96.2 (American Type Culture Collection), ST88-14 (a gift from Dr. Jonathan Fletcher, Brigham and Women's Hospital), and T265 (a gift from Dr. George De Vries, Hines VA Hospital) and the non-NF1 sporadic human MPNST cell line STS26T (a gift from Dr. Steven Porcelli, Albert Einstein College of Medicine) were used for our studies and maintained and propagated as described previously (22).

Techniques: Western Blot, Expressing, Cell Culture

Journal: Scientific Reports

Article Title: Circulating MicroRNA-92b-3p as a Novel Biomarker for Monitoring of Synovial Sarcoma

doi: 10.1038/s41598-017-12660-5

Figure Lengend Snippet: Differential diagnosis of synovial sarcoma from other soft tissue sarcomas by circulating/secreted miR-92b-3p . ( A , B ) The expression of miR-92b-3p and miR-150-3p in cells ( A ) and cell culture media ( B ) of SS (SYO-1, YaFuSS, HS-SY-II, and Yamato-SS) and the other STS cell lines (HT1080, NMS2, and UPS2023). The Mann-Whitney U test was used for comparison between groups. ( C ) Serum miR-92b-3p levels in SS patients and other STS patients ( p < 0.001). The Mann-Whitney U test was used for comparison between groups. ( D ) ROC curve analysis of miR-92b-3p discriminating SS patients from other STS patients. AUC was 0.88 (95% confidence interval = 0.72–1.0). ( E ) Serum miR-150-3p expression levels in SS patients and other STS patients. The Mann-Whitney U test was used for comparison between groups. ( F ) ROC curve analysis of miR-150-3p discriminating SS patients from other STS patients. AUC was 0.54 (95% confidence interval = 0.33–0.75).

Article Snippet: A human malignant peripheral nerve sheath tumour cell line NMS2 was available from the cell bank of RIKEN BioResource Center (Ibaraki, Japan).

Techniques: Biomarker Discovery, Expressing, Cell Culture, MANN-WHITNEY, Comparison

Journal: Clinical Cancer Research

Article Title: MEK Inhibition Synergizes with TYK2 Inhibitors in NF1-Associated Malignant Peripheral Nerve Sheath Tumors

doi: 10.1158/1078-0432.CCR-22-3722

Figure Lengend Snippet: TYK2 inhibitors reduce proliferation in MPNST cells. A, TYK2 protein levels were visualized by IHC in MPNST and plexiform neurofibroma, with positive staining scored on a 0 to 3 scale. B, Relative TYK2 protein levels in MPNST cell lines by WES Western blot analysis. Protein bands analyzed by densitometry, with TYK2 normalized to β-actin. C, JW23.3 and JH-2–002 cells were treated with TYK2 inhibitors (WU-12, WU-76) or inactive control (WU-18) for 3 days, and cell confluence was determined by IncuCyte assay. D, Representative images of IncuCyte assay at 72 hours, with YOYO-1 green fluorescence as indicator of apoptosis.

Article Snippet: Human-derived MPNST cell lines were generated by the Pratilas lab at Johns Hopkins University (JH-2–002 cells), and obtained through the Johns Hopkins NF1 biospecimen repository , or obtained from the Fletcher lab at Dana Farber Cancer Institute (MPNST-724 cells, RRID:CVCL_AU20; ref. ).

Techniques: Staining, WES Western Blot, Control, Fluorescence

Journal: Clinical Cancer Research

Article Title: MEK Inhibition Synergizes with TYK2 Inhibitors in NF1-Associated Malignant Peripheral Nerve Sheath Tumors

doi: 10.1158/1078-0432.CCR-22-3722

Figure Lengend Snippet: TYK2 inhibition leads to compensatory stimulation of the MAPK pathway in MPNST cells. A, JH-2–002 cells were treated with 40 μmol/L WU-12 or control for 48 hours, and gene expression was analyzed by qPCR array for JAK/STAT pathway–related genes. Log 2 fold changes of significantly changed genes are graphed. B, JW23.3 cells were treated with control or 40 μmol/L WU-12 or WU-76 for 48 hours. Global gene expression was determined by RNA-seq pathway analysis. Changes in gene expression for WU-12 versus control were examined by RNA-seq for genes downstream of STAT3 in a heatmap ( C ) and boxplot ( D ), as well as MEK/MAPK pathway genes in a volcano plot ( E ), enrichment plot ( F ), and boxplot ( G ). P < 0.05 vs. vehicle control.

Article Snippet: Human-derived MPNST cell lines were generated by the Pratilas lab at Johns Hopkins University (JH-2–002 cells), and obtained through the Johns Hopkins NF1 biospecimen repository , or obtained from the Fletcher lab at Dana Farber Cancer Institute (MPNST-724 cells, RRID:CVCL_AU20; ref. ).

Techniques: Inhibition, Control, Gene Expression, RNA Sequencing

Journal: Clinical Cancer Research

Article Title: MEK Inhibition Synergizes with TYK2 Inhibitors in NF1-Associated Malignant Peripheral Nerve Sheath Tumors

doi: 10.1158/1078-0432.CCR-22-3722

Figure Lengend Snippet: TYK2 inhibitory drugs decrease activation of STAT3 while increasing activation of ERK1/2 in MPNST cells. A, JW23.3 cells and ( B ) JH-2–002 cells were incubated with a TYK2 inhibitor (40 μmol/L WU-12) or vehicle control for the indicated times. Phosphorylated and total protein levels for STAT3 and ERK1/2 were analyzed by the WES Western system. Bands were analyzed by densitometry in the WES software, with phosphorylated protein normalized to the matching total protein and expressed as percent of control at the same time point. *, P < 0.05 vs. vehicle control at the same time point.

Article Snippet: Human-derived MPNST cell lines were generated by the Pratilas lab at Johns Hopkins University (JH-2–002 cells), and obtained through the Johns Hopkins NF1 biospecimen repository , or obtained from the Fletcher lab at Dana Farber Cancer Institute (MPNST-724 cells, RRID:CVCL_AU20; ref. ).

Techniques: Activation Assay, Incubation, Control, Western Blot, Software

Journal: Clinical Cancer Research

Article Title: MEK Inhibition Synergizes with TYK2 Inhibitors in NF1-Associated Malignant Peripheral Nerve Sheath Tumors

doi: 10.1158/1078-0432.CCR-22-3722

Figure Lengend Snippet: The specific TYK2 inhibitor deucravacitinib (BMS-986165) decreases MPNST cell proliferation at lower doses. The MPNST cell lines JW23.3 ( A and B ), JH-2–002 ( C and D ), and MPNST-724 ( E and F ) were treated with the indicated doses of the specific TYK2 inhibitor deucravacitinib or the pan-JAK inhibitor baricitinib for 3 days in IncuCyte live cell proliferation assays.

Article Snippet: Human-derived MPNST cell lines were generated by the Pratilas lab at Johns Hopkins University (JH-2–002 cells), and obtained through the Johns Hopkins NF1 biospecimen repository , or obtained from the Fletcher lab at Dana Farber Cancer Institute (MPNST-724 cells, RRID:CVCL_AU20; ref. ).

Techniques:

Journal: Clinical Cancer Research

Article Title: MEK Inhibition Synergizes with TYK2 Inhibitors in NF1-Associated Malignant Peripheral Nerve Sheath Tumors

doi: 10.1158/1078-0432.CCR-22-3722

Figure Lengend Snippet: Inhibitors of TYK2 (deucravacitinib) and MEK (mirdametinib) act synergistically to reduce proliferation and increase apoptosis in JW23.3 MPNST cells. Cell confluence and apoptosis were analyzed by the IncuCyte assay after 48-hour incubation with drugs. Synergy was analyzed using Synergy Finder software by the HSA method for ( A ) inhibition of cell proliferation or ( B ) apoptosis, and mean synergy score is reported ( P < 0.05 vs. drug alone). Intensity of red color indicates synergy score for each dose combination, whereas green indicates antagonism. C, Representative images of JW23.3 cells treated for 48 hours. YOYO-1 green fluorescence indicates apoptotic cells.

Article Snippet: Human-derived MPNST cell lines were generated by the Pratilas lab at Johns Hopkins University (JH-2–002 cells), and obtained through the Johns Hopkins NF1 biospecimen repository , or obtained from the Fletcher lab at Dana Farber Cancer Institute (MPNST-724 cells, RRID:CVCL_AU20; ref. ).

Techniques: Incubation, Software, Inhibition, Fluorescence

Journal: Clinical Cancer Research

Article Title: MEK Inhibition Synergizes with TYK2 Inhibitors in NF1-Associated Malignant Peripheral Nerve Sheath Tumors

doi: 10.1158/1078-0432.CCR-22-3722

Figure Lengend Snippet: The combination of drugs inhibiting TYK2 and MEK block MPNST tumor growth in mice. A, Schematic diagram of treatment paradigm. Mice with JW23.3 MPNST xenograft tumors ( n = 6 per group; B ), WU-386 MPNST PDX tumors ( n = 3 per group; C ), or JH-2–002 MPNST xenograft tumors ( n = 5 per group; D ) were treated daily with 1.5 mg/kg mirdametinib (Mirda), 30 mg/kg deucravacitinib (Deucra, BMS-986165), the combination of drugs, or vehicle control for 3 weeks or until tumors reached the maximum allowed volume. *, P < 0.05 vs. vehicle control; a , P < 0.05 for drug combination vs. drugs alone. E and F, Diagram of TYK2/STAT3 and MEK/ERK pathways after treatment with Deucra and/or Mirda in MPNST cells. (Illustrations were created with BioRender.com .)

Article Snippet: Human-derived MPNST cell lines were generated by the Pratilas lab at Johns Hopkins University (JH-2–002 cells), and obtained through the Johns Hopkins NF1 biospecimen repository , or obtained from the Fletcher lab at Dana Farber Cancer Institute (MPNST-724 cells, RRID:CVCL_AU20; ref. ).

Techniques: Blocking Assay, Control

Journal: bioRxiv

Article Title: PRC2 Inactivating Mutations in Cancer are Synthetic Lethal with DNMT1 Targeted Therapy via Enhanced Viral Mimicry

doi: 10.1101/2022.05.28.493429

Figure Lengend Snippet: A-B , Dose-response curves and IC 50 of a pan-DNMT inhibitor, decitabine (DAC), or a DNMT1-selective inhibitor (GSK862) in SUZ12 -isogenic MPNST cells: PRC2-wt= M3-sg Con ; PRC2-loss = M3-sg SUZ12 . C , Western blot analysis of SUZ12- isogenic cells treated with 3 μM EPZ-6438 (an EZH2 inhibitor) or 5 μM EED226 (an EED inhibitor). D , Dose-response curves of GSK862 in SUZ12- isogenic cells treated with EZH2 or EED inhibitors, or vehicle controls. E , Western blot validation and DNMT inhibitor (DAC and GSK862) dose-response curves for Eed- isogenic murine MPNST cells derived from Nf1 -/- ; Cdkn2a -/- ; Cdkn2b -/- genetically engineered murine models. F , Dose-response curves of DAC and GSK862 in SUZ12 -null human MPNSTs cells with or without PRC2 ( SUZ12 ) restoration. G-H , Growth curves over time of orthotopically transplanted human PRC2-wt (M3) and PRC2-loss (M14) cell-line-derived xenografts (CDXs) in CB-17 SCID mice treated with DAC or vehicle control by luciferase bioluminescence imaging. I , Fold change tumor volume of subcutaneous PRC2-loss human MPNST patient derived xenograft (PDX-1) treated with 5 mg/kg DAC or vehicle control. J , Luciferase bioluminescence-based growth curve over time of orthotopically transplanted human PRC2-loss M14 CDX in SCID mice treated with 45 mg/kg GSK3685032 or vehicle. All error bars: mean ± SEM; Student’s two-tailed T-test (** P <0.01; *** P <0.001; **** P <0.0001).

Article Snippet: For DAC studies on human MPNST xenograft bearing CB-17 SCID mice (Taconic): all cells were implanted into mice 1:1 with Matrigel (BD Biosciences).

Techniques: Western Blot, Biomarker Discovery, Derivative Assay, Control, Luciferase, Imaging, Two Tailed Test

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Retooling a Blood-based Biomarker: Phase I Assessment of the High-Affinity CA19-9 Antibody HuMab-5B1 for Immuno-PET Imaging of Pancreatic Cancer

doi: 10.1158/1078-0432.CCR-18-3667

Figure Lengend Snippet: A, There is a reduction in liver uptake of radioactivity with increasing antibody mass (error bars show one standard deviation). B, Maximum-intensity projection image demonstrates the effect in a patient injected with 3 mg antibody (left) and a (different) patient injected with 47 mg HuMab 5B1 antibody 2 h before injection of 3 mg radiolabeled HuMab 5B (right). C, Fused PET/CT, PET, and contrast-enhanced CT of PDAC liver metastases in a patient who received 47 mg of HuMab 5B1 before injection of the radiolabeled antibody. Small liver metastases measuring less than 5 mm (red arrows) are visualized with high contrast. SUVs values were 101.4, 70.7, and 53.4 g/ml, respectively. Arrowhead indicates splenic infarct.

Article Snippet: The fully human HuMab-5B1 antibody was manufactured for MabVax Therapeutics Holdings, Inc. (San Diego, CA) by Patheon Biologics (Groningen, Netherlands) under current good manufacturing practice (cGMP) guidelines and supplied in sterile vials to MSK.

Techniques: Radioactivity, Standard Deviation, Injection, Positron Emission Tomography-Computed Tomography