Journal: Investigational new drugs

Article Title: Preclinical characterization of atiprimod, a novel JAK2 AND JAK3 inhibitor

doi: 10.1007/s10637-010-9429-z

Figure Lengend Snippet: Atiprimod inhibits the phosphorylation of JAK2, and JAK3. FDCP-EpoR cell lines carrying JAK2WT or JAK2V617F, human SET2 and CMK cells were treated with equipotent doses (IC20, IC50, and IC80) of atiprimod for 4 h. Cells were lysed and whole lysates were immunoprecipitated with anti-JAK2 and AK3 antibodies, (a) Atiprimod inhibits the phosphorylation of JAK2 in FDCP-EpoR cell lines carrying constitutively active JAK2V617F(a) and the phosphorylation of JAK2 in human SET2 cells and JAK3 in CMK cells (b)

Article Snippet: The human ET (at megakaryoblastic leukemic transformation) cell line carrying the JAK2 V617F mutation (SET-2, ACC608) and the mutant JAK3-positive human acute megakaryocytic leukemia carrying the mutation A572V within the JAK3 pseudokinase (CMK, ACC393) were purchased from DSMZ (Braunschweig, Germany) and maintained in RPMI 1,640, supplemented with 20% (SET-2) or 10% FBS (CMK).

Techniques: Phospho-proteomics, Immunoprecipitation

Journal: Investigational new drugs

Article Title: Preclinical characterization of atiprimod, a novel JAK2 AND JAK3 inhibitor

doi: 10.1007/s10637-010-9429-z

Figure Lengend Snippet: Induction of apoptosis (A) and mitochondrial potential damage (B) by atiprimod at equipotent concentrations (IC 50 and IC 80 ) in FDCP-EpoR cell lines transduced with either JAK2 WT or JAK2 V617F

Article Snippet: The human ET (at megakaryoblastic leukemic transformation) cell line carrying the JAK2 V617F mutation (SET-2, ACC608) and the mutant JAK3-positive human acute megakaryocytic leukemia carrying the mutation A572V within the JAK3 pseudokinase (CMK, ACC393) were purchased from DSMZ (Braunschweig, Germany) and maintained in RPMI 1,640, supplemented with 20% (SET-2) or 10% FBS (CMK).

Techniques: Transduction, Membrane

Journal: Investigational new drugs

Article Title: Preclinical characterization of atiprimod, a novel JAK2 AND JAK3 inhibitor

doi: 10.1007/s10637-010-9429-z

Figure Lengend Snippet: Quantitation of the JAK2V617F mutant allele and effect of atiprimod on JAK2V617F allele burden. Peripheral blood low-density cells from 3 patients with PV were incubated in the presence of atripimod at doses ranging from 0.25 to 2.0 µM in clonogenic assays and evaluated after 14 days. Ten colonies at each dose were microaspirated and the JAK2V617F allele burden was quantitated by qRT-PCR. After DNA extraction, gDNA was used for PCR amplification with JAK2-exonl4 primers. Quantitative allele-specific suppressive PCR (ASS-PCR) was performed using the purified PCR product on a sequence detection system 7,000 platform

Article Snippet: The human ET (at megakaryoblastic leukemic transformation) cell line carrying the JAK2 V617F mutation (SET-2, ACC608) and the mutant JAK3-positive human acute megakaryocytic leukemia carrying the mutation A572V within the JAK3 pseudokinase (CMK, ACC393) were purchased from DSMZ (Braunschweig, Germany) and maintained in RPMI 1,640, supplemented with 20% (SET-2) or 10% FBS (CMK).

Techniques: Quantitation Assay, Mutagenesis, Incubation, Quantitative RT-PCR, DNA Extraction, Amplification, Purification, Sequencing

Journal: Molecular and Clinical Oncology

Article Title: Analysis of genes encoding epigenetic regulators in myeloproliferative neoplasms: Coexistence of a novel SETBP1 mutation in a patient with a p.V617F JAK2 positive myelofibrosis

doi: 10.3892/mco.2019.1840

Figure Lengend Snippet: Human cancer cell lines tested.

Article Snippet: Informed consent was obtained from all patients and procedures were approved by the Ethical Committee on Clinical Research of University of Navarra. table ft1 table-wrap mode="anchored" t5 Table II. caption a7 Cell line name Repository (number) Origin A-549 DSMZ (ACC-107) Lung carcinoma BK-006 ECACC p.V617F JAK2 positive PV BK-013 ECACC (98100924) p.V617F JAK2 negative ET BK015 ECACC (99092421) p.V617F JAK2 negative ET DAUDI DSMZ (ACC-78) Burkitt lymphoma EOL-1 DSMZ (ACC-386) Acute myeloid eosinophilic leukemia F-36P DSMZ (ACC-543) Acute myeloid leukemia secondary to myelodysplastic syndrome (MDS) HCC-1937 DSMZ (ACC-513) Breast carcinoma HEL DSMZ ACC-11 Erythroleukemia HL60 DSMZ ACC-3 Acute myeloid leukemia (AML) HU3 Dr Morgan, USA Acute megakaryoblastic leukemia K-562 DSMZ ACC-10 BCR-ABL1 positive chronic myeloid leukemia (CML) in blast crisis KARPAS-299 DSMZ ACC-31 T cell lymphoma KARPAS-422 DSMZ ACC-32 B cell lymphoma M-07e DSMZ ACC-104 Acute megakaryoblastic leukemia MG-63 ATCC: CRL-1427 Osteosarcoma MOLM-13 DSMZ ACC-554 Acute myeloid leukemia (AML) MOLT-16 DSMZ ACC-29 T cell acute lymphoid leukemia (T-ALL) MOLT-4 DSMZ ACC-362 Acute lymphoid leukemia (ALL) MV-411 DSMZ ACC-102 Acute monoblastic/monocytic leukemia RAJI DSMZ ACC-319 Burkitt lymphoma REH DSMZ ACC-22 B-ALL (t(12;21) (p13;q22) (fusion ETV6-RUNX1 )) SET2 DSMZ ACC-608 p.V617F JAK2 positive ET TF-1 DSMZ ACC-334 Erythroleukemia UKE-1 Dr W. Fiedelr, Hamburg, Germany p.V617F JAK2 positive ET transformed to AML Open in a separate window ECACC, European Collection of Cell Culture; DSMZ, Deutsche Sammlung von Mikroorganismen und Zellkulturen.

Techniques: Transformation Assay

Journal: Leukemia research

Article Title: Clinical and biological relevance of CREB3L1 in Philadelphia chromosome-negative myeloproliferative neoplasms.

doi: 10.1016/j.leukres.2022.106883

Figure Lengend Snippet: Fig. 1. Clinical relevance of CREB3L1 expres sion in platelets in myeloproliferative neoplasms (MPNs). Box plot comparing CREB3L1 mRNA expression levels determined by reverse- transcription quantitative polymerase chain re action (RT-qPCR) between granulocytes (Gran), lymphocytes (Lym), red blood cells (RBC) and platelets (A), CREB3L1 expression levels in platelet RNA between MPN cases harboring CALR mutation (CALRm) and JAK2 V617F (JAK2VF) (B), CREB3L1 expression levels in platelet RNA among patients with essential thrombocytopenia (ET) grouped by age (< 60 or ≥60 years at the time of sample collection) (C), conventional risk score (D), IPSET (E), and r- IPSET risk score (F). Kaplan–Meier probability curves for overall survival (G), myelofibrosis- free survival (H), and event-free survival (I) in patients with ET expressing lower (first quartile) or higher (fourth quartile) CREB3L1 mRNA levels in platelets, determined by RT-qPCR.

Article Snippet: HEL cells and SET-2 cells harboring endogenous JAK2 V617F [24], and UT-7/TPO/CALRm cells exogenously expressing either CALR Del52 or Ins5 mutations, previously generated by retroviral transfection of pMSCV-IRES-mOrange vector (Addgene #54568) containing CALRm sequences [23,25], were then virally infected at an efficiency of ~20%, as previously described [26].

Techniques: Expressing, Reverse Transcription, Quantitative RT-PCR, Mutagenesis

Journal: Leukemia research

Article Title: Clinical and biological relevance of CREB3L1 in Philadelphia chromosome-negative myeloproliferative neoplasms.

doi: 10.1016/j.leukres.2022.106883

Figure Lengend Snippet: Fig. 3. Downregulation of UPR genes expression in platelets of patients with essen tial thrombocytopenia (ET) and patients with polycythemia vera (PV). Box plot comparing PERK (A), ATF6 (B), IRE1 (C) and CREB3L1 (D) mRNA expression levels determined by reverse-transcription quantitative polymerase chain reaction in platelet RNA of patients with ET grouped by driver mutations (CALR mu tation; CALRm, and JAK2 V617F; JAK2VF) and those of patients with PV (PERK (E), ATF6 (F), IRE1 (G), and CREB3L1 (H)) relative to healthy controls.

Article Snippet: HEL cells and SET-2 cells harboring endogenous JAK2 V617F [24], and UT-7/TPO/CALRm cells exogenously expressing either CALR Del52 or Ins5 mutations, previously generated by retroviral transfection of pMSCV-IRES-mOrange vector (Addgene #54568) containing CALRm sequences [23,25], were then virally infected at an efficiency of ~20%, as previously described [26].

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Investigational new drugs

Article Title: Preclinical characterization of atiprimod, a novel JAK2 AND JAK3 inhibitor

doi: 10.1007/s10637-010-9429-z

Figure Lengend Snippet: Atiprimod inhibits the phosphorylation of JAK2, and JAK3. FDCP-EpoR cell lines carrying JAK2WT or JAK2V617F, human SET2 and CMK cells were treated with equipotent doses (IC20, IC50, and IC80) of atiprimod for 4 h. Cells were lysed and whole lysates were immunoprecipitated with anti-JAK2 and AK3 antibodies, (a) Atiprimod inhibits the phosphorylation of JAK2 in FDCP-EpoR cell lines carrying constitutively active JAK2V617F(a) and the phosphorylation of JAK2 in human SET2 cells and JAK3 in CMK cells (b)

Article Snippet: The human ET (at megakaryoblastic leukemic transformation) cell line carrying the JAK2 V617F mutation (SET-2, ACC608) and the mutant JAK3-positive human acute megakaryocytic leukemia carrying the mutation A572V within the JAK3 pseudokinase (CMK, ACC393) were purchased from DSMZ (Braunschweig, Germany) and maintained in RPMI 1,640, supplemented with 20% (SET-2) or 10% FBS (CMK).

Techniques: Phospho-proteomics, Immunoprecipitation

Journal: Molecular and Clinical Oncology

Article Title: Analysis of genes encoding epigenetic regulators in myeloproliferative neoplasms: Coexistence of a novel SETBP1 mutation in a patient with a p.V617F JAK2 positive myelofibrosis

doi: 10.3892/mco.2019.1840

Figure Lengend Snippet: Human cancer cell lines tested.

Article Snippet: Informed consent was obtained from all patients and procedures were approved by the Ethical Committee on Clinical Research of University of Navarra. table ft1 table-wrap mode="anchored" t5 Table II. caption a7 Cell line name Repository (number) Origin A-549 DSMZ (ACC-107) Lung carcinoma BK-006 ECACC p.V617F JAK2 positive PV BK-013 ECACC (98100924) p.V617F JAK2 negative ET BK015 ECACC (99092421) p.V617F JAK2 negative ET DAUDI DSMZ (ACC-78) Burkitt lymphoma EOL-1 DSMZ (ACC-386) Acute myeloid eosinophilic leukemia F-36P DSMZ (ACC-543) Acute myeloid leukemia secondary to myelodysplastic syndrome (MDS) HCC-1937 DSMZ (ACC-513) Breast carcinoma HEL DSMZ ACC-11 Erythroleukemia HL60 DSMZ ACC-3 Acute myeloid leukemia (AML) HU3 Dr Morgan, USA Acute megakaryoblastic leukemia K-562 DSMZ ACC-10 BCR-ABL1 positive chronic myeloid leukemia (CML) in blast crisis KARPAS-299 DSMZ ACC-31 T cell lymphoma KARPAS-422 DSMZ ACC-32 B cell lymphoma M-07e DSMZ ACC-104 Acute megakaryoblastic leukemia MG-63 ATCC: CRL-1427 Osteosarcoma MOLM-13 DSMZ ACC-554 Acute myeloid leukemia (AML) MOLT-16 DSMZ ACC-29 T cell acute lymphoid leukemia (T-ALL) MOLT-4 DSMZ ACC-362 Acute lymphoid leukemia (ALL) MV-411 DSMZ ACC-102 Acute monoblastic/monocytic leukemia RAJI DSMZ ACC-319 Burkitt lymphoma REH DSMZ ACC-22 B-ALL (t(12;21) (p13;q22) (fusion ETV6-RUNX1 )) SET2 DSMZ ACC-608 p.V617F JAK2 positive ET TF-1 DSMZ ACC-334 Erythroleukemia UKE-1 Dr W. Fiedelr, Hamburg, Germany p.V617F JAK2 positive ET transformed to AML Open in a separate window ECACC, European Collection of Cell Culture; DSMZ, Deutsche Sammlung von Mikroorganismen und Zellkulturen.

Techniques: Transformation Assay