1   87 151 A PHASE 1 STUDY OF AZACITIDINE WITH HIGH-DOSE CYTARABINE AND MITOXANTRONE IN HIGH-RISK ACUTE MYELOID LEUKEMIA. IN THIS PHASE 1 STUDY, AZACITIDINE (AZA) WAS GIVEN BEFORE HIGH-DOSE CYTARABINE (HIDAC) AND MITOXANTRONE (MITO) BASED ON THE HYPOTHESIS THAT EPIGENETIC PRIMING WITH A HYPOMETHYLATING AGENT BEFORE CYTOTOXIC CHEMOTHERAPY WOULD IMPROVE RESPONSE RATES IN PATIENTS WITH HIGH-RISK ACUTE MYELOID LEUKEMIA (AML), INCLUDING RELAPSED/REFRACTORY DISEASE. THE PRIMARY OBJECTIVE WAS TO ESTABLISH THE RECOMMENDED PHASE 2 DOSE OF AZA GIVEN BEFORE STANDARD HIDAC/MITO. IN A DOSE ESCALATION SCHEME, 46 PATIENTS (MEDIAN AGE, 66 YEARS) RECEIVED AZA AT 37.5, 50, OR 75 MG/M2 SUBCUTANEOUSLY OR IV ONCE DAILY ON DAYS 1 TO 5 FOLLOWED BY HIDAC (3000 MG/M2) AND MITOXANTRONE (30 MG/M2) ONCE EACH ON DAYS 6 AND 10 (THE HIDAC/MITO DOSE WAS REDUCED 33% IN ELDERLY SUBJECTS). TWO DOSE-LIMITING TOXICITIES OCCURRED (BOTH IN THE SAME PATIENT): ACUTE LIVER FAILURE AND KIDNEY INJURY AT THE 50 MG/M2 DOSE. THE 30-DAY INDUCTION DEATH RATE WAS 2.2% (1 OF 46). THE OVERALL RESPONSE RATE, INCLUDING COMPLETE REMISSION AND COMPLETE REMISSION WITH INCOMPLETE COUNT RECOVERY, WAS 61% (28 OF 46). PREVIOUSLY UNTREATED PATIENTS AGED >/=60 YEARS WITH THERAPY-RELATED AML AND DE NOVO AML WERE MORE LIKELY TO RESPOND THAN UNTREATED PATIENTS WITH AML PROGRESSING FROM AN ANTECEDENT HEMATOLOGIC DISORDER (MYELODYSPLASTIC SYNDROME AND CHRONIC MYELOMONOCYTIC LEUKEMIA). PATIENTS WITH FAVORABLE EUROPEAN LEUKEMIA NETWORK RISK (P = .008), NPM1 MUTATIONS (P = .007), OR IDH2 MUTATIONS (P = .03) WERE MORE LIKELY TO RESPOND, AND THOSE WITH TP53 MUTATIONS (P = .03) WERE LESS LIKELY TO RESPOND. THE RECOMMENDED PHASE 2 DOSE OF AZA IS 75 MG/M2 PER DAY ON DAYS 1 TO 5 FOLLOWED BY HIDAC (3000 MG/M2) AND MITOXANTRONE (30 MG/M2) ONCE EACH ON DAYS 6 AND 10. THIS TRIAL WAS REGISTERED AT WWW.CLINICALTRIALS.GOV AS #NCT01839240.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
2   88  43 A PHASE I BIOLOGICAL STUDY OF MG98, AN OLIGODEOXYNUCLEOTIDE ANTISENSE TO DNA METHYLTRANSFERASE 1, IN PATIENTS WITH HIGH-RISK MYELODYSPLASIA AND ACUTE MYELOID LEUKEMIA. PURPOSE: EPIGENETIC SILENCING VIA ABERRANT PROMOTER DNA HYPERMETHYLATION OF NORMAL GENES HAS BEEN DESCRIBED AS A LEUKEMOGENIC MECHANISM IN MYELODYSPLASTIC SYNDROMES (MDS) AND ACUTE MYELOID LEUKEMIAS (AML). WE HYPOTHESIZED THAT MG98, AN OLIGONUCLEOTIDE ANTISENSE TO DNA METHYLTRANSFERASE 1 (DNMT1), COULD REVERSE MALIGNANT PHENOTYPES BY DOWN-REGULATING DNMT1 AND INDUCING REEXPRESSION OF HYPERMETHYLATED GENES. THIS PHASE I STUDY WAS CONDUCTED TO DETERMINE A BIOLOGICALLY EFFECTIVE DOSE AND DESCRIBE THE SAFETY OF MG98 IN MDS/AML. EXPERIMENTAL DESIGN: TWENTY-THREE PATIENTS WITH MDS (N = 11) AND AML (N = 12) WERE ENROLLED. BIOLOGICALLY EFFECTIVE DOSE WAS DEFINED AS THE DOSE AT WHICH > OR =50% OF PATIENTS EXPERIENCED >50% REDUCTION IN DNMT1 EXPRESSION WITH ACCEPTABLE TOXICITY. ESCALATING DOSES OF MG98 WERE ADMINISTERED ACCORDING TO TWO SCHEDULES (2-HOUR I.V. BOLUS FOLLOWED BY 5-DAY CONTINUOUS I.V. INFUSION EVERY 14 DAYS, OR 14-DAY CONTINUOUS I.V. INFUSION EVERY 21 DAYS). RESULTS: DNMT1 DOWN-REGULATION WAS OBSERVED IN 8 PATIENTS. HOWEVER, BIOLOGICALLY EFFECTIVE DOSE WAS NOT REACHED. REEXPRESSION OF TARGET GENES (P15, WIT1, AND ER) WAS OBSERVED IN 12 PATIENTS BUT DID NOT CORRELATE WITH DNMT1 DOWN-REGULATION. ESCALATION WAS STOPPED DUE TO DOSE-LIMITING TOXICITIES (BONE PAIN, NAUSEA, AND FEVER). NO OBJECTIVE CLINICAL RESPONSE WAS OBSERVED. DISEASE STABILIZATION OCCURRED IN 6 (26%) PATIENTS. CONCLUSIONS: NO PHARMACODYNAMIC OR CLINICAL ACTIVITY WAS OBSERVED AT MG98 DOSES AND SCHEDULES ADMINISTERED. DESPITE THIS, PURSUING DNMT1 DOWN-REGULATION REMAINS A SOUND APPROACH FOR TARGETING ABERRANT EPIGENETICS IN AML/MDS. FUTURE STUDIES WITH DIFFERENT FORMULATION AND/OR DOSES AND SCHEDULES WILL BE REQUIRED TO ENSURE EFFICIENT MG98 INTRACELLULAR UPTAKE AND FULLY EVALUATE ITS THERAPEUTIC POTENTIAL.	2008	
                                                                                                                                                                                                                                                                                                                                                                                                   
3 5056  30 PHASE I TRIAL OF LOW DOSE DECITABINE TARGETING DNA HYPERMETHYLATION IN PATIENTS WITH CHRONIC LYMPHOCYTIC LEUKAEMIA AND NON-HODGKIN LYMPHOMA: DOSE-LIMITING MYELOSUPPRESSION WITHOUT EVIDENCE OF DNA HYPOMETHYLATION. TARGETING ABERRANT DNA HYPERMETHYLATION IN CHRONIC LYMPHOCYTIC LEUKAEMIA (CLL) AND NON-HODGKIN LYMPHOMA (NHL) WITH DECITABINE MAY REVERSE EPIGENETIC SILENCING IN B-CELL MALIGNANCIES. TWENTY PATIENTS WERE ENROLLED IN TWO PHASE I TRIALS TO DETERMINE THE MINIMUM EFFECTIVE PHARMACOLOGICAL DOSE OF DECITABINE IN PATIENTS WITH RELAPSED/REFRACTORY CLL (N = 16) AND NHL (N = 4). PATIENTS RECEIVED 1-3 CYCLES OF DECITABINE. DOSE-LIMITING TOXICITY (DLT) WAS OBSERVED IN 2 OF 4 CLL AND 2 OF 2 NHL PATIENTS RECEIVING DECITABINE AT 15 MG/M(2) PER D DAYS 1-10, CONSISTING OF GRADE 3-4 THROMBOCYTOPENIA AND HYPERBILIRUBINAEMIA. SIX PATIENTS WITH CLL RECEIVED DECITABINE AT 10 MG/M(2) PER D DAYS 1-10 WITHOUT DLT; HOWEVER, RE-EXPRESSION OF METHYLATED GENES OR CHANGES IN GLOBAL DNA METHYLATION WERE NOT OBSERVED. THEREFORE, A 5-DAY DECITABINE SCHEDULE WAS EXAMINED. WITH 15 MG/M(2) PER D DECITABINE DAYS 1-5, DLT OCCURRED IN 2 OF 6 CLL AND 2 OF 2 NHL PATIENTS, CONSISTING OF GRADE 3-4 NEUTROPENIA, THROMBOCYTOPENIA, AND FEBRILE NEUTROPENIA. EIGHT PATIENTS HAD STABLE DISEASE. IN 17 PATIENTS, THERE WERE NO SIGNIFICANT CHANGES IN GENOME-WIDE METHYLATION OR IN TARGET GENE RE-EXPRESSION. IN CONCLUSION, DOSE-LIMITING MYELOSUPPRESSION AND INFECTIOUS COMPLICATIONS PREVENTED DOSE ESCALATION OF DECITABINE TO LEVELS ASSOCIATED WITH CHANGES IN GLOBAL METHYLATION OR GENE RE-EXPRESSION IN CLL AND NHL.	2010	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
4 2464  45 EPIGENETIC THERAPY USING BELINOSTAT FOR PATIENTS WITH UNRESECTABLE HEPATOCELLULAR CARCINOMA: A MULTICENTER PHASE I/II STUDY WITH BIOMARKER AND PHARMACOKINETIC ANALYSIS OF TUMORS FROM PATIENTS IN THE MAYO PHASE II CONSORTIUM AND THE CANCER THERAPEUTICS RESEARCH GROUP. PURPOSE: EPIGENETIC ABERRATIONS HAVE BEEN REPORTED IN HEPATOCELLULAR CARCINOMA (HCC). IN THIS STUDY OF PATIENTS WITH UNRESECTABLE HCC AND CHRONIC LIVER DISEASE, EPIGENETIC THERAPY WITH THE HISTONE DEACETYLASE INHIBITOR BELINOSTAT WAS ASSESSED. THE OBJECTIVES WERE TO DETERMINE DOSE-LIMITING TOXICITY AND MAXIMUM-TOLERATED DOSE (MTD), TO ASSESS PHARMACOKINETICS IN PHASE I, AND TO ASSESS ACTIVITY OF AND EXPLORE POTENTIAL BIOMARKERS FOR RESPONSE IN PHASE II. PATIENTS AND METHODS: MAJOR ELIGIBILITY CRITERIA INCLUDED HISTOLOGICALLY CONFIRMED UNRESECTABLE HCC, EUROPEAN COOPERATIVE ONCOLOGY GROUP PERFORMANCE SCORE </= 2, AND ADEQUATE ORGAN FUNCTION. PHASE I CONSISTED OF 18 PATIENTS; BELINOSTAT WAS GIVEN INTRAVENOUSLY ONCE PER DAY ON DAYS 1 TO 5 EVERY 3 WEEKS; DOSE LEVELS WERE 600 MG/M(2) PER DAY (LEVEL 1), 900 MG/M(2) PER DAY (LEVEL 2), 1,200 MG/M(2) PER DAY (LEVEL 3), AND 1,400 MG/M(2) PER DAY (LEVEL 4). PHASE II CONSISTED OF 42 PATIENTS. THE PRIMARY END POINT WAS PROGRESSION-FREE SURVIVAL (PFS), AND THE MAIN SECONDARY END POINTS WERE RESPONSE ACCORDING TO RESPONSE EVALUATION CRITERIA IN SOLID TUMORS (RECIST) AND OVERALL SURVIVAL (OS). EXPLORATORY ANALYSIS WAS CONDUCTED ON PRETREATMENT TUMOR TISSUES TO DETERMINE WHETHER HR23B EXPRESSION IS A POTENTIAL BIOMARKER FOR RESPONSE. RESULTS: BELINOSTAT PHARMACOKINETICS WERE LINEAR FROM 600 TO 1,400 MG/M(2) WITHOUT SIGNIFICANT ACCUMULATION. THE MTD WAS NOT REACHED AT THE MAXIMUM DOSE ADMINISTERED. DOSE LEVEL 4 WAS USED IN PHASE II. THE MEDIAN NUMBER OF CYCLES WAS TWO (RANGE, ONE TO 12). THE PARTIAL RESPONSE (PR) AND STABLE DISEASE (SD) RATES WERE 2.4% AND 45.2%, RESPECTIVELY. THE MEDIAN PFS AND OS WERE 2.64 AND 6.60 MONTHS, RESPECTIVELY. EXPLORATORY ANALYSIS REVEALED THAT DISEASE STABILIZATION RATE (COMPLETE RESPONSE PLUS PR PLUS SD) IN TUMORS HAVING HIGH AND LOW HR23B HISTOSCORES WERE 58% AND 14%, RESPECTIVELY (P = .036). CONCLUSION: EPIGENETIC THERAPY WITH BELINOSTAT DEMONSTRATES TUMOR STABILIZATION AND IS GENERALLY WELL-TOLERATED. HR23B EXPRESSION WAS ASSOCIATED WITH DISEASE STABILIZATION.	2012	
                             
5 2465  36 EPIGENETIC THERAPY WITH HYDRALAZINE AND MAGNESIUM VALPROATE REVERSES IMATINIB RESISTANCE IN PATIENTS WITH CHRONIC MYELOID LEUKEMIA. THE EPIGENETIC DRUGS HYDRALAZINE AND VALPROATE WERE ADMINISTERED IN A COMPASSIONATE MANNER TO 8 PATIENTS WITH CHRONIC MYELOID LEUKEMIA (CML) REFRACTORY TO IMATINIB. TWO PATIENTS HAD A COMPLETE HEMATOLOGIC RESPONSE (25%),1 MAJOR CYTOGENETIC RESPONSE, 1 COMPLETE CYTOGENETIC RESPONSE (25% ANY CYTOGENETIC RESPONSE), AND 3 (37.5%)STABLE DISEASE. NO GRADE 3 OR 4 TOXICITY WAS OBSERVED. THESE RESULTS SHOW THE ABILITY OF EPIGENETIC THERAPY TO REVERT IMATINIB RESISTANCE. BACKGROUND: EPIGENETIC ALTERATIONS PARTICIPATE IN THE DEVELOPMENT OF ACQUIRED RESISTANCE TO IMATINIB, HENCE, THE DNA METHYLATION, AND HISTONE DEACETYLASE INHIBITORS HYDRALAZINE AND VALPROATE, RESPECTIVELY, HAS THE POTENTIAL TO OVERCOME IT. PATIENT AND METHODS: A SERIES OF 8 PATIENTS WITH CHRONIC MYELOID LEUKEMIA (CML) REFRACTORY TO IMATINIB MESYLATE WITH NO ACCESS TO SECOND-GENERATION TYROSINE KINASE INHIBITORS WERE TREATED WITH HYDRALAZINE AND VALPROATE IN A COMPASSIONATE MANNER. CLINICAL EFFICACY AND SAFETY OF THESE DRUGS ADDED TO IMATINIB MESYLATE WERE EVALUATED. RESULTS: TWO PATIENTS WERE IN THE BLAST PHASE, 5 WERE IN THE ACCELERATED PHASE, AND 1 WAS IN THE CHRONIC PHASE. ALL THE PATIENTS CONTINUED WITH THE SAME DOSE OF IMATINIB THAT THEY HAD BEEN RECEIVING AT THE TIME OF DEVELOPMENT OF RESISTANCE, WITH A MEDIAN DOSE OF 600 MG DAILY (RANGE, 400-800 MG). THE MEDIAN TIME FROM DIAGNOSIS OF CML TO THE START OF HYDRALAZINE AND VALPROATE WAS 53.6 MONTHS (RANGE, 19-84 MONTHS). TWO (25%) PATIENTS HAD A COMPLETE HEMATOLOGIC RESPONSE, ONE (12.5%) HAD AN MAJOR CYTOGENETIC RESPONSE, AND ONE (12.5%) HAD A COMPLETE CYTOGENETIC RESPONSE. THREE (37.5%) PATIENTS HAD STABLE DISEASE, AND ONLY ONE (12.5%) PATIENT FAILED TO RESPOND. AT A MEDIAN FOLLOW-UP TIME OF 18 MONTHS (RANGE, 3-18 MONTHS), THE MEDIAN SURVIVAL HAD NOT BEEN REACHED, AND THE PROJECTED OVERALL SURVIVAL WAS 63%. ALL THE PATIENTS HAD MILD NEUROLOGIC TOXICITY, INCLUDING DISTAL TREMOR AND SOMNOLENCE. NO GRADE 3 OR 4 TOXICITY WAS OBSERVED. CONCLUSIONS: OUR RESULTS SUGGEST THAT THE EPIGENETIC DRUGS HYDRALAZINE AND VALPROATE REVERT THE RESISTANCE TO IMATINIB IN PATIENTS WITH CML.	2012	
                                                                                                              
6 6689  41 VALPROIC ACID AT THERAPEUTIC PLASMA LEVELS MAY INCREASE 5-AZACYTIDINE EFFICACY IN HIGHER RISK MYELODYSPLASTIC SYNDROMES. PURPOSE: EPIGENETIC CHANGES PLAY A ROLE AND COOPERATE WITH GENETIC ALTERATIONS IN THE PATHOGENESIS OF MYELODYSPLASTIC SYNDROMES (MDS). WE CONDUCTED A PHASE II MULTICENTER STUDY ON THE COMBINATION OF THE DNA-METHYLTRANSFERASE INHIBITOR 5-AZACYTIDINE (5-AZA) AND THE HISTONE DEACETYLASE INHIBITOR VALPROIC ACID (VPA) IN PATIENTS WITH HIGHER RISK MDS. EXPERIMENTAL DESIGN: WE ENROLLED 62 PATIENTS WITH MDS (REFRACTORY ANEMIA WITH EXCESS BLASTS, 39 PATIENTS; REFRACTORY ANEMIA WITH EXCESS BLASTS IN TRANSFORMATION, 19 PATIENTS; AND CHRONIC MYELOMANOCYTIC LEUKEMIA (CMML), 4 PATIENTS) AND AN INTERNATIONAL PROGNOSTIC SCORING SYSTEM (IPSS) RATING OF INTERMEDIATE-2 (42 PATIENTS) OR HIGH (20 PATIENTS). VPA WAS GIVEN TO REACH A PLASMA CONCENTRATION OF >50 MICROG/ML, THEN 5-AZA WAS ADDED S.C. AT 75 MG/M(2) FOR 7 DAYS IN EIGHT MONTHLY CYCLES. RESULTS: THE MEDIAN OVERALL SURVIVAL WAS 14.4 MONTHS. AT A MEDIAN FOLLOW-UP OF 12 MONTHS (RANGE, 0.7-21.0), THE DISEASE PROGRESSED IN 20 PATIENTS, WITH 21% CUMULATIVE INCIDENCE OF PROGRESSION. OF 26 PATIENTS WHO COMPLETED EIGHT CYCLES, 30.7% OBTAINED COMPLETE OR PARTIAL REMISSION, 15.4% HAD A MAJOR HEMATOLOGIC IMPROVEMENT, WHEREAS 38.5% SHOWED STABLE DISEASE. DRUG-RELATED TOXICITY WAS MILD. FAVORABLE PROGNOSTIC FACTORS FOR SURVIVAL WERE IPSS INTERMEDIATE-2 AND PLASMA VPA OF > OR =50 MICROG/ML (LOG RANK = 0.013 AND 0.007, RESPECTIVELY). ANALYSIS OF POLYMORPHISMS IMPORTANT FOR THE METABOLISM OF THE DRUGS USED IN THE TRIAL SHOWED THAT CARRIERS OF THE CYP2C19*2 VARIANT OF CYTOCHROME P450 REQUIRED HIGHER VPA DOSES TO ACHIEVE THE TARGET VPA PLASMA CONCENTRATION OF 50 MICROG/ML ON DAY 1 OF 5-AZA TREATMENT (P = 0.0021). CONCLUSION: OUR DATA SHOW THAT THE 5-AZA/VPA COMBINATION IS ACTIVE AND SAFE IN PATIENTS WITH MDS WITH A POOR PROGNOSIS. ACHIEVEMENT OF VPA THERAPEUTIC LEVELS MAY INDEED INCREASE 5-AZA EFFICACY.	2009	
                                                                                                                                                                                                                                                                                                                                                                                                
7 5478  45 RESULTS OF A RANDOMIZED STUDY OF 3 SCHEDULES OF LOW-DOSE DECITABINE IN HIGHER-RISK MYELODYSPLASTIC SYNDROME AND CHRONIC MYELOMONOCYTIC LEUKEMIA. EPIGENETIC THERAPY WITH HYPOMETHYLATING DRUGS IS NOW THE STANDARD OF CARE IN MYELODYSPLASTIC SYNDROME (MDS). RESPONSE RATES REMAIN LOW, AND MECHANISM-BASED DOSE OPTIMIZATION HAS NOT BEEN REPORTED. WE INVESTIGATED THE CLINICAL AND PHARMACODYNAMIC RESULTS OF DIFFERENT DOSE SCHEDULES OF DECITABINE. ADULTS WITH ADVANCED MDS OR CHRONIC MYELOMONOCYTIC LEUKEMIA (CMML) WERE RANDOMIZED TO 1 OF 3 DECITABINE SCHEDULES: (1) 20 MG/M2 INTRAVENOUSLY DAILY FOR 5 DAYS; (2) 20 MG/M2 SUBCUTANEOUSLY DAILY FOR 5 DAYS; AND (3) 10 MG/M2 INTRAVENOUSLY DAILY FOR 10 DAYS. RANDOMIZATION FOLLOWED A BAYESIAN ADAPTIVE DESIGN. NINETY-FIVE PATIENTS WERE TREATED (77 WITH MDS, AND 18 WITH CMML). OVERALL, 32 PATIENTS (34%) ACHIEVED A COMPLETE RESPONSE (CR), AND 69 (73%) HAD AN OBJECTIVE RESPONSE BY THE NEW MODIFIED INTERNATIONAL WORKING GROUP CRITERIA. THE 5-DAY INTRAVENOUS SCHEDULE, WHICH HAD THE HIGHEST DOSE-INTENSITY, WAS SELECTED AS OPTIMAL; THE CR RATE IN THAT ARM WAS 39%, COMPARED WITH 21% IN THE 5-DAY SUBCUTANEOUS ARM AND 24% IN THE 10-DAY INTRAVENOUS ARM (P < .05). THE HIGH DOSE-INTENSITY ARM WAS ALSO SUPERIOR AT INDUCING HYPOMETHYLATION AT DAY 5 AND AT ACTIVATING P15 EXPRESSION AT DAYS 12 OR 28 AFTER THERAPY. WE CONCLUDE THAT A LOW-DOSE, DOSE-INTENSITY SCHEDULE OF DECITABINE OPTIMIZES EPIGENETIC MODULATION AND CLINICAL RESPONSES IN MDS.	2007	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
8 5665  33 SF3B1, RUNX1 AND TP53 MUTATIONS SIGNIFICANTLY IMPACT THE OUTCOME OF PATIENTS WITH LOWER-RISK MYELODYSPLASTIC SYNDROME. INTRODUCTION: PROGNOSIS OF PATIENTS WITH MYELODYSPLASTIC SYNDROME (MDS), PARTICULARLY THE GROUP WITH LOWER-RISK DISEASE (LR-MDS) IS VERY HETEROGENEOUS. SEVERAL STUDIES HAVE DESCRIBED THE PROGNOSTIC VALUE OF RECURRENT SOMATIC MUTATIONS IN MDS INCLUDING ALL RISK CATEGORIES. RECENTLY, THE INCORPORATION OF GENOMIC DATA TO CLINICAL PARAMETERS DEFINED THE NEW MOLECULAR INTERNATIONAL PROGNOSTIC SCORING SYSTEM (IPSS-M). MATERIALS AND METHODS: IN THIS STUDY, WE EVALUATED THE IMPACT OF MOLECULAR PROFILE IN A SERIES OF 181 PATIENTS WITH LR-MDS AND NON-PROLIFERATIVE CHRONIC MYELOMONOCYTIC LEUKEMIA. RESULTS: EPIGENETIC REGULATORS (TET2, ASXL1) AND SPLICING (SF3B1) WERE THE MOST RECURRENT MUTATED PATHWAYS. IN UNIVARIATE ANALYSIS, RUNX1 OR TP53 MUTATIONS CORRELATED WITH LOWER MEDIAN OVERALL SURVIVAL (OS). IN CONTRAST, SF3B1 MUTATION WAS ASSOCIATED WITH PROLONGED MEDIAN OS [95 MONTHS (95% IC, 32-157) VS. 33 MONTHS (95% CI, 19-46) IN UNMUTATED PATIENTS (P < 0.01)]. IN A MULTIVARIATE COX REGRESSION MODEL, RUNX1 MUTATIONS INDEPENDENTLY ASSOCIATED WITH SHORTER OS, WHILE SF3B1 MUTATION RETAINED ITS FAVORABLE IMPACT ON OUTCOME (HR: 0.24, 95% CI, 0.1-0.5; P = 0.001). IN ADDITION, TP53 OR RUNX1 MUTATIONS WERE IDENTIFIED AS PREDICTIVE COVARIATES FOR THE PROBABILITY OF LEUKEMIC PROGRESSION (P < 0.001). CONCLUSION: INCORPORATION OF MOLECULAR TESTING IN LR-MDS IDENTIFIED A SUBSET OF PATIENTS WITH EXPECTED POORER OUTCOME, EITHER DUE TO LOWER SURVIVAL OR PROBABILITY OF LEUKEMIC PROGRESSION.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
9 3560  34 IMPACT OF CLINICAL, CYTOGENETIC, AND MOLECULAR PROFILES ON LONG-TERM SURVIVAL AFTER TRANSPLANTATION IN PATIENTS WITH CHRONIC MYELOMONOCYTIC LEUKEMIA. CHRONIC MYELOMONOCYTIC LEUKEMIA (CMML) IS A HETEROGENEOUS GROUP OF CLONAL HEMATOPOIETIC MALIGNANCIES WITH VARIABLE CLINICAL AND MOLECULAR FEATURES. WE ANALYZED LONG-TERM RESULTS OF ALLOGENEIC HEMATOPOIETIC CELL TRANSPLANTATION IN PATIENTS WITH CMML AND DETERMINED CLINICAL AND MOLECULAR RISK FACTORS ASSOCIATED WITH OUTCOMES. DATA FROM 129 PATIENTS, AGED 7-74 (MEDIAN 55) YEARS, AT VARIOUS STAGES OF THE DISEASE AND TRANSPLANTED FROM RELATED OR UNRELATED DONORS WERE ANALYZED. USING A PANEL OF 75 GENES SOMATIC MUTATIONS PRESENT BEFORE HEMATOPOIETIC CELL TRANSPLANTATION WERE IDENTIFIED IN 52 PATIENTS. THE PROGRESSION-FREE SURVIVAL RATE AT 10 YEARS WAS 29%. THE MAJOR CAUSE OF DEATH WAS RELAPSE (32%), WHICH WAS SIGNIFICANTLY ASSOCIATED WITH ADVERSE CYTOGENETICS (HAZARD RATIO, 3.77; P=0.0002), CMML PROGNOSTIC SCORING SYSTEM (HAZARD RATIO, 14.3, P=0.01), AND MD ANDERSON PROGNOSTIC SCORES (HAZARD RATIO, 9.4; P=0.005). MORTALITY WAS ASSOCIATED WITH HIGH-RISK CYTOGENETICS (HAZARD RATIO, 1.88; P=0.01) AND HIGH HEMATOPOIETIC CELL TRANSPLANTATION COMORBIDITY INDEX (SCORE >/=4: HAZARD RATIO, 1.99; P=0.01). HIGH OVERALL MUTATION BURDEN (>/=10 MUTATIONS: HAZARD RATIO, 3.4; P=0.02), AND >/=4 MUTATED EPIGENETIC REGULATORY GENES (HAZARD RATIO 5.4; P=0.003) WERE LINKED TO RELAPSE. UNSUPERVISED CLUSTERING OF THE CORRELATION MATRIX REVEALED DISTINCT HIGH-RISK GROUPS WITH UNIQUE ASSOCIATIONS OF MUTATIONS AND CLINICAL FEATURES. CMML WITH A HIGH MUTATION BURDEN APPEARED TO BE DISTINCT FROM HIGH-RISK GROUPS DEFINED BY COMPLEX CYTOGENETICS. NEW TRANSPLANT STRATEGIES MUST BE DEVELOPED TO TARGET SPECIFIC DISEASE SUBGROUPS, STRATIFIED BY MOLECULAR PROFILING AND CLINICAL RISK FACTORS.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
10 5984  36 TET2, DNMT3A, IDH1, AND JAK2 MUTATION IN MYELOPROLIFERATIVE NEOPLASMS IN SOUTHERN IRAN. BACKGROUND: FIVE EPIGENETIC REGULATOR MUTATIONS ARE CONSIDERED IN MYELOPROLIFERATIVE NEOPLASMS (MPN) THAT HAVE PROGNOSTIC AND THERAPEUTIC VALUES. OBJECTIVE: WE AIMED TO EVALUATE THESE MUTATIONS IN MPNS AMONG THE IRANIAN POPULATION. METHODS: WE SELECTED 5 MUTATIONS IN 4 EPIGENETIC REGULATORY GENES [TET2, DNMT3A, IDH1 (RS147001633&RS121913499), AND JAK2)] AND EVALUATED 130 PATIENTS WITH MPNS INCLUDING 78 PHILADELPHIA CHROMOSOME NEGATIVE (49 ETS, 20 PVS, AND 9 PMFS) AND 52 PHILADELPHIA CHROMOSOME-POSITIVE PATIENTS AS WELL AS 51 HEALTHY CONTROLS. RESULTS: EIGHT PATIENTS (6.5%) CARRIED THE DNMT3A MUTATION, 35 (27%) WERE POSITIVE FOR TET2 MUTATION AND 64 (49.3%) HAD THE JAK2V617F MUTATION. IN THE HEALTHY CONTROLS, 16 (31.4%) CASES HAD THE TET2 MUTATION (15 HETEROZYGOTE + 1 HOMOZYGOTE) AND ONE HAD HETEROZYGOTE JAK2 MUTATION. THERE WAS NO STATISTICALLY SIGNIFICANT DIFFERENCE BETWEEN PATIENT GROUPS FOR ANY OF THESE MUTATIONS, EXCEPT FOR JAK2. THE JAK2 MUTATION RATE WAS 18 (90%), 25 (51%), 7 (77.8%), 14 (26.9%) IN POLYCYTHEMIA VERA, ESSENTIAL THROMBOCYTHEMIA, PRIMARY MYELOFIBROSIS, AND CHRONIC MYELOCYTIC LEUKEMIA, RESPECTIVELY. PATIENTS AGED 60 AND OLDER WERE MORE LIKELY TO CARRY THE TET2 MUTATION (23% VS. 39% IN YOUNGER AND OLDER THAN 60 YEARS OLD INDIVIDUALS, P=0.025). IDH1 WAS NOT DETECTED AT ALL AND PV HAD THE HIGHEST TET2 MUTATION 7(35%). TWO PMF PATIENTS HAD A HISTORY OF BONE MARROW TRANSPLANTATION THAT WERE NEGATIVE FOR IDH1AND DNMT3A AND ONE WAS POSITIVE FOR TET2 MUTATION. CONCLUSION: IN THE NORMAL IRANIAN POPULATION, THE HETEROZYGOTE FORM OF TET2 MUTATION IS SIGNIFICANT, ESPECIALLY IN THE ELDERLY. NO ASSOCIATION WAS FOUND BETWEEN JAK2 AND TET2 MUTATIONS. BOTH OF THEM ARE MORE PREVALENT IN THE AGE GROUP OF 60 YEARS AND OLDER. DNMT3A MUTATION HAS A LOW PREVALENCE AND OCCURS IN BOTH POSITIVE AND NEGATIVE MPNS.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                
11 4571  26 MYELOMONOCYTIC SKEWING IN CHRONIC MYELOMONOCYTIC LEUKEMIA: PHENOTYPIC, MOLECULAR AND BIOLOGIC FEATURES AND IMPACT ON SURVIVAL. BACKGROUND: MYELOMONOCYTIC SKEWING IS CONSIDERED AS A KEY PATHOPHYSIOLOGIC PHENOMENON IN CHRONIC MYELOMONOCYTIC LEUKEMIA (CMML), BUT ITS PREVALENCE AND POTENTIAL CORRELATION WITH PHENOTYPIC, GENOTYPIC, AND CLINICAL FEATURES ARE POORLY DEFINED. METHODS: SKEWED DIFFERENTIATION TOWARD THE MYELOMONOCYTIC OVER ERYTHROID COMMITMENT AS INDICATED BY AN INVERSE RATIO OF MYELOMONOCYTIC/ERYTHROID COLONIES WAS INVESTIGATED IN 146 PATIENTS WITH CMML BY SEMISOLID IN VITRO CULTURES. RESULTS: THERE WAS A HIGH PREVALENCE OF MYELOMONOCYTIC SKEWING IN PATIENTS WITH CMML (120/146, 82%); WHEREAS, THIS PHENOMENON WAS RARE IN NORMAL INDIVIDUALS (1/98, 1%). PATIENTS WITH CMML WITH MYELOMONOCYTIC SKEWING HAD HIGHER WHITE BLOOD CELL AND PERIPHERAL BLAST CELL COUNTS, AND LOWER PLATELET VALUES. THE NUMBER OF MUTATIONS IN GENES OF THE EPIGENETIC AND/OR SPLICING CATEGORY WAS HIGHER IN CMML PATIENTS WITH AS COMPARED WITH PATIENTS WITHOUT SKEWING. PATIENTS WITH MYELOMONOCYTIC SKEWING HAD MORE FREQUENTLY MUTATIONS IN RASOPATHY GENES AND HIGHER GROWTH FACTOR INDEPENDENT MYELOID COLONY FORMATION. INTERESTINGLY, THE LACK OF MYELOMONOCYTIC SKEWING DISCRIMINATED PATIENTS WITH CMML WITH A PARTICULARLY FAVORABLE PROGNOSIS (60 VS 19 MONTHS, P = .003) AND A MINIMAL RISK OF TRANSFORMATION. CONCLUSION: MYELOMONOCYTIC SKEWING AS DETERMINED BY SEMISOLID CULTURES CAN DISCRIMINATE SUBGROUPS OF PATIENTS WITH CMML WITH A DIFFERENT PHENOTYPE, A DIFFERENT GENOTYPE, AND A DIFFERENT PROGNOSIS.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
12 5283  44 PROPHYLACTIC OR PREEMPTIVE LOW-DOSE AZACITIDINE AND DONOR LYMPHOCYTE INFUSION TO PREVENT DISEASE RELAPSE FOLLOWING ALLOGENEIC TRANSPLANTATION IN PATIENTS WITH HIGH-RISK ACUTE MYELOGENOUS LEUKEMIA OR MYELODYSPLASTIC SYNDROME. BECAUSE OF THE PERSISTENTLY HIGH RATES OF RELAPSE OF PATIENTS WITH HIGH-RISK ACUTE MYELOGENOUS LEUKEMIA (AML) AND MYELODYSPLASTIC SYNDROME (MDS) FOLLOWING ALLOGENEIC HEMATOPOIETIC STEM CELL TRANSPLANTATION (ALLO-HSCT), POST-TRANSPLANTATION MAINTENANCE THERAPY HAS BEEN PROPOSED. WE PREVIOUSLY INITIATED A PHASE II TRIAL IN WHICH EPIGENETIC THERAPY WAS COMBINED WITH IMMUNOTHERAPY IN AN ATTEMPT TO REDUCE DISEASE RELAPSE. IN THAT STUDY, LOW-DOSE AZACITIDINE (AZA) AND ESCALATING DOSES OF DONOR LYMPHOCYTE INFUSION (DLI) WERE GIVEN AS POST-ALLO-HSCT MAINTENANCE TREATMENT. IN THE PRESENT STUDY, WE RETROSPECTIVELY ANALYZE A LARGER COHORT OF PATIENTS RECEIVING POST-TRANSPLANTATION MAINTENANCE THERAPY AND PROVIDE UPDATES ON SOME PATIENTS OF THE EARLIER STUDY. THE OBJECTIVES OF THE PRESENT STUDY WERE TO ANALYZE THE CUMULATIVE INCIDENCE OF RELAPSE (CIR), OVERALL SURVIVAL (OS), AND PROGRESSION-FREE SURVIVAL (PFS) AND THE INCIDENCE OF ACUTE AND CHRONIC GRAFT-VERSUS-HOST DISEASE (GVHD) OF PATIENTS WITH HIGH-RISK AML OR MDS RECEIVING POST-TRANSPLANTATION MAINTENANCE TREATMENT WITH AZA WITH OR WITHOUT DLI. WE RETROSPECTIVELY ANALYZED 77 PATIENTS (54 WITH AML, 23 WITH MDS) CONSIDERED AT HIGH RISK BASED ON EITHER THEIR GENOMIC OR CLINICAL STATUS AT TRANSPLANTATION. FOLLOWING ALLOGENEIC TRANSPLANTATION, THEY RECEIVED AT LEAST 1 CYCLE OF PROPHYLACTIC OR PREEMPTIVE LOW-DOSE AZA WITH OR WITHOUT ESCALATING DOSES OF DLI TO PREVENT DISEASE RELAPSE. ALMOST ONE-HALF OF THE PATIENTS (47%) WERE ABLE TO RECEIVE THE FULL 12 CYCLES OF SCHEDULED AZA, AND A MAJORITY (79%) RECEIVED AT LEAST 1 DLI. WITH A MEDIAN FOLLOW-UP OF 24 MONTHS, 19 PATIENTS (25%; 16 WITH AML, 3 WITH MDS) RELAPSED, AT A MEDIAN OF 9.8 MONTHS (RANGE, 4 TO 58.6 MONTHS), GIVING A 22% CIR AT 24 MONTHS. OS AND PFS AT 24 MONTHS WERE 70.8% AND 68.3%, RESPECTIVELY. THE CUMULATIVE INCIDENCES OF GRADE II-IV ACUTE GVHD AND CHRONIC GVHD WERE 27.4% AND 45%, RESPECTIVELY. ONLY A MINORITY OF PATIENTS (11%) REQUIRED DELAYED ADMINISTRATION OF AZA. THESE FINDINGS CONFIRM THAT AZA-DLI MAINTENANCE IS BOTH TOLERABLE AND EFFECTIVE IN REDUCING THE RISK OF POST-TRANSPLANTATION RELAPSE.	2021	

13 4540  27 MULTISTEP PATHOGENESIS OF CHRONIC MYELOMONOCYTIC LEUKEMIA IN PATIENTS. BACKGROUND: A MULTISTEP PATHOGENESIS OF MYELOID LEUKEMIA INCLUDING MUTATIONS IN EPIGENETIC, SPLICEOSOME, AND SIGNALING GENES HAS BEEN RECENTLY DEMONSTRATED IN A PRECLINICAL MODEL BUT IS POORLY VALIDATED IN PATIENTS. METHODS: CLINICAL, PHENOTYPIC, AND BIOLOGIC FEATURES WERE COMPARED BETWEEN THREE DISTINCT MOLECULARLY DEFINED CMML COHORTS INCLUDING TET2 MONOMUTATED PATIENTS (T, N = 10), TET2/SRSF2 BIMUTATED PATIENTS (TS, N = 19), AND PATIENTS WHO HAD NRAS MUTATIONS IN ADDITION TO TET2/SRSF2 COMUTATIONS (TSN, N = 14). RESULTS: MEDIAN SURVIVAL WAS 90, 45, AND 9 MONTHS, RESPECTIVELY (P = .001). WHEREAS NO PATIENT IN THE T AND TS GROUP TRANSFORMED INTO ACUTE MYELOID LEUKEMIA (AML), 6/14 PATIENTS IN THE TSN GROUP HAD AML AT STUDY ENTRY OR TRANSFORMED DURING FOLLOW-UP. LEUKOCYTE COUNTS, BLAST CELL COUNTS, AND LDH LEVELS WERE SIGNIFICANTLY HIGHER IN TSN VS. TS AND T, RESPECTIVELY, WHEREAS HEMOGLOBIN AND PLATELET VALUES WERE NOT SIGNIFICANTLY DIFFERENT. INCREASED GROWTH FACTOR-INDEPENDENT MYELOID COLONY FORMATION WAS RESTRICTED TO TSN BUT NOT FOUND IN T AND TS, RESPECTIVELY. THE PROPORTION OF PATIENTS SHOWING IN VITRO MYELOMONOCYTIC SKEWING IN T, TS, AND TSN WAS 0%, 56%, AND 100%, RESPECTIVELY (P = .010). CONCLUSION: OUR RESULTS DEMONSTRATE THAT THE MODEL OF MULTISTEP PATHOGENESIS IN CMML CAN BE RECAPITULATED IN PATIENTS REGARDING CLINICAL, PHENOTYPIC, AND BIOLOGIC FEATURES.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
14 2822  43 FIRST-IN-HUMAN STUDY OF INHALED AZACITIDINE IN PATIENTS WITH ADVANCED NON-SMALL CELL LUNG CANCER. BACKGROUND: AEROSOLIZED AZACITIDINE HAS BEEN SHOWN TO INHIBIT ORTHOTOPIC LUNG CANCER GROWTH AND INDUCE RE-EXPRESSION OF METHYLATED TUMOR SUPPRESSOR GENES IN MURINE MODELS. WE HYPOTHESIZED THAT INHALED AZACITIDINE IS SAFE AND EFFECTIVE IN REVERSING EPIGENETIC CHANGES IN THE BRONCHIAL EPITHELIUM SECONDARY TO CHRONIC SMOKING. PATIENTS AND METHODS: WE REPORT THE FIRST IN HUMAN STUDY OF INHALED AZACITIDINE. AZACITIDINE IN AQUEOUS SOLUTION WAS USED TO GENERATE AN AEROSOL SUSPENSION OF 0.25-5 MUM PARTICLE SIZE. MAIN INCLUSION CRITERIA: STAGE IV OR RECURRENT NSCLC WITH PREDOMINANTLY LUNG INVOLVEMENT, >/=1 PRIOR SYSTEMIC THERAPY, ECOG PS 0-1, AND ADEQUATE PULMONARY FUNCTION. PATIENTS RECEIVED INHALED AZACITIDINE DAILY ON DAYS 1-5 AND 15-19 OF 28-DAY CYCLES, AT 3 ESCALATING DOSES (15, 30 AND 45 MG/M(2) DAILY). THE PRIMARY OBJECTIVE WAS TO DETERMINE THE FEASIBILITY AND TOLERABILITY OF THIS NEW THERAPEUTIC MODALITY. THE KEY SECONDARY OBJECTIVES INCLUDED PHARMACOKINETICS, METHYLATION PROFILES AND EFFICACY. RESULTS: FROM 3/2015 TO 2/2018, EIGHT PATIENTS RECEIVED A MEDIAN NUMBER OF 2 (IQR = 1) CYCLES OF INHALED AZACITIDINE. NO CLINICALLY SIGNIFICANT ADVERSE EVENTS WERE OBSERVED, EXCEPT ONE PATIENT TREATED AT THE HIGHEST DOSE DEVELOPED AN ASYMPTOMATIC GRADE 2 DECREASED DLCO WHICH RESOLVED SPONTANEOUSLY. ONE PATIENT RECEIVING 12 CYCLES OF THERAPY HAD AN OBJECTIVE AND DURABLE PARTIAL RESPONSE, AND TWO PATIENTS HAD STABLE DISEASE. PLASMA AZACITIDINE WAS ONLY BRIEFLY DETECTABLE IN PATIENTS TREATED AT THE HIGHER DOSES. MOREOVER, IN 2 OF 3 PARTICIPANTS WHO AGREED AND UNDERWENT PRE- AND POST-TREATMENT BRONCHOSCOPY, THE GLOBAL DNA METHYLATION IN THE BRONCHIAL EPITHELIUM DECREASED BY 24 % AND 79 % POST-THERAPY, RESPECTIVELY. THE INTERVAL BETWEEN LAST INHALED TREATMENT AND BRONCHOSCOPY WAS 3 DAYS. CONCLUSIONS: INHALED AZACITIDINE RESULTED IN NEGLIGIBLE PLASMA LEVELS COMPARED TO THE PREVIOUSLY REPORTED SUBCUTANEOUS ADMINISTRATION AND WAS WELL-TOLERATED. THE RESULTS JUSTIFY THE CONTINUED DEVELOPMENT OF INHALED AZACITIDINE AT NON-CYTOTOXIC DOSES FOR PATIENTS WITH LUNG-CONFINED MALIGNANT AND/OR PREMALIGNANT LESIONS.	2021	
                                                                                                                                        
15 2769  53 EXTENDED DOSING WITH CC-486 (ORAL AZACITIDINE) IN PATIENTS WITH MYELOID MALIGNANCIES. CC-486 (ORAL AZACITIDINE) IS AN EPIGENETIC MODIFIER IN CLINICAL DEVELOPMENT FOR TREATMENT OF HEMATOLOGICAL CANCERS. THIS STUDY OF EXTENDED CC-486 DOSING INCLUDED PATIENTS WITH MYELODYSPLASTIC SYNDROMES (MDSS), CHRONIC MYELOMONOCYTIC LEUKEMIA (CMML), OR ACUTE MYELOID LEUKEMIA (AML). AFTER A PHARMACOKINETIC ASSESSMENT PERIOD, 31 PATIENTS (MDS N = 18, CMML N = 4, AND AML N = 9) ENTERED A CLINICAL PHASE IN WHICH THEY RECEIVED CC-486 300 MG ONCE-DAILY FOR 21 DAYS OF REPEATED 28-DAY CYCLES. MEDIAN AGE WAS 71 YEARS (RANGE: 53-93); 42% OF PATIENTS WERE AGED >/=75 YEARS. A TOTAL OF 5 PATIENTS WITH AML (63%) HAD PRIOR MDS. MEDIAN NUMBER OF CC-486 TREATMENT CYCLES WAS 4 (RANGE: 1-32). THE MOST COMMON TREATMENT-EMERGENT ADVERSE EVENTS (TEAES) WERE GASTROINTESTINAL (84% OF PATIENTS) AND HEMATOLOGIC (81%). MOST COMMON GRADE 3-4 TEAES WERE NEUTROPENIA (N = 13, 42%) AND ANEMIA (N = 9, 29%). TEN PATIENTS EXPERIENCED GRADE 4 NEUTROPENIA. INFREQUENTLY, CC-486 DOSE WAS INTERRUPTED OR REDUCED DUE TO GASTROINTESTINAL (N = 5, 16%) OR HEMATOLOGIC (N = 6, 19%) TEAES. OVERALL RESPONSE RATE (COMPLETE REMISSION [CR], CR WITH INCOMPLETE HEMATOLOGICAL RECOVERY [CRI], PARTIAL REMISSION [PR], MARROW CR) IN THE MDS/CMML SUBGROUPS WAS 32% AND IN THE AML SUBGROUP (CR/CRI/PR) WAS 22%. RED BLOOD CELL TRANSFUSION INDEPENDENCE RATES IN THE MDS/CMML AND AML SUBGROUPS WERE 33% AND 25%, RESPECTIVELY, AND 2 MDS/CMML PATIENTS ATTAINED HEMATOLOGIC IMPROVEMENT AS A BEST RESPONSE ON-STUDY. NO BASELINE GENE MUTATION WAS PREDICTIVE OF RESPONSE/NONRESPONSE. CC-486 ALLOWS FLEXIBLE DOSING AND SCHEDULES TO IMPROVE TOLERABILITY OR RESPONSE. NEUTROPENIA IN EARLY TREATMENT CYCLES DESERVES SCRUTINY AND MAY WARRANT INITIATION OF PROPHYLACTIC ANTIBIOTICS. KEY POINTS: THE SAFETY PROFILE OF ORAL CC-486 WAS COMPARABLE TO THAT OF INJECTABLE AZACITIDINE; MOST ADVERSE EVENTS WERE HEMATOLOGICAL AND GASTROINTESTINAL. EXTENDED (21-DAY/CYCLE) CC-486 DOSING INDUCED RESPONSES IN PATIENTS WITH HEMATOLOGICAL MALIGNANCIES, MANY OF WHOM HAD PRIOR DNMTI FAILURE.	2018	
                                                                                                                                                                                                                                                         
16  831  30 CHARACTERIZATION OF TET AND IDH GENE EXPRESSION IN CHRONIC LYMPHOCYTIC LEUKEMIA: COMPARISON WITH NORMAL B CELLS AND PROGNOSTIC SIGNIFICANCE. BACKGROUND: CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) IS THE MOST COMMON HEMATOLOGICAL MALIGNANCY IN WESTERN COUNTRIES, CHARACTERIZED BY A HETEROGENEOUS CLINICAL COURSE. ALTHOUGH GENETIC STUDIES HAVE IDENTIFIED CHROMOSOMAL ABERRATIONS OR SPECIFIC MUTATIONS, EPIGENETIC CHANGES HAVE BEEN POORLY CHARACTERIZED IN CLL. METHODS: WE ASSESSED TEN-ELEVEN TRANSLOCATIONS (TET) 1, 2, AND 3, ISOCITRATE DEHYDROGENASE (IDH) 1, AND 2 MESSENGER RNA (MRNA) EXPRESSION USING REAL-TIME PCR ON PURIFIED LEUKEMIC B CELLS FROM 214 CLL PATIENTS (MEDIAN FOLLOW-UP = 75 MONTHS, RANGE 1-380), NORMAL PERIPHERAL BLOOD B CELLS (N = 20), AND UMBILICAL CORD BLOOD B CELLS (N = 21). THE MICROENVIRONMENT INFLUENCE WAS ASSESSED AFTER 24 H CO-CULTURE OF CLL CELLS WITH BONE MARROW MESENCHYMAL STROMAL CELLS (BMSC). FINALLY, 5-HYDROXYMETHYLCYTOSINE LEVEL (%5-HMC) WAS ASSESSED BY ELISA IN CLL CELLS ALONE OR WITH MICROENVIRONMENT STIMULI. RESULTS: TET 1 AND 3 AND IDH2 WERE DECREASED IN CLL CELLS COMPARED WITH HEALTHY B CELLS (P = 0.0221, 0.0013, <0.0001, RESPECTIVELY), WHILE IDH1 WAS OVEREXPRESSED (P = 0.0037). TET2 AND IDH1 WERE SIGNIFICANTLY CORRELATED WITH TREATMENT-FREE SURVIVAL (TFS); PATIENTS WITH HIGH TET2/IDH1 EXPRESSION HAD A HIGHER MEDIAN TFS (111 MONTHS) THAN PATIENTS WITH LOW EXPRESSION (78 MONTHS, P = 0.0071/0.0123). MOREOVER, TET1 EXPRESSION DECREASED (P = 0.0371), WHILE TET3 AND IDH2 EXPRESSION INCREASED (P = 0.0273/0.0039) IN CO-CULTURES. HOWEVER, %5-HMC WAS NOT CORRELATED WITH CLINICAL DATA AND WAS UNCHANGED FOLLOWING MICROENVIRONMENT STIMULI. CONCLUSIONS: DESPITE A SLIGHT DEREGULATION IN CLL CELLS COMPARED WITH NORMAL B CELLS, WE IDENTIFIED A SIGNIFICANT ASSOCIATION BETWEEN TET/IDH GENE EXPRESSION AND PROGNOSIS, SUGGESTING THAT EPIGENETIC CHANGES COULD POTENTIALLY BE ASSOCIATED WITH DISEASE PROGRESSION. MOREOVER, DESPITE AN IDENTICAL %5-HMC, TET GENE EXPRESSION WAS INFLUENCED BY CONTACT WITH BMSC CONFIRMING THE CRUCIAL ROLE OF THE MICROENVIRONMENT IN CLL PATHOGENESIS.	2016	
                                                                                                                                                                                                                                                        
17 3168  33 GTPASE REGULATOR ASSOCIATED WITH THE FOCAL ADHESION KINASE (GRAF) TRANSCRIPT WAS DOWN-REGULATED IN PATIENTS WITH MYELOID MALIGNANCIES. BACKGROUND: GTPASE REGULATOR ASSOCIATED WITH THE FOCAL ADHESION KINASE (GRAF), A PUTATIVE TUMOR SUPPRESSOR GENE, IS FOUND INACTIVATED IN HEMATOPOIETIC MALIGNANCIES BY EITHER GENETIC OR EPIGENETIC ABNORMALITIES. HOWEVER, THE EXPRESSION LEVEL OF GRAF GENE HAS NOT YET BEEN STUDIED IN LEUKEMIA. THE AIM OF THIS STUDY WAS TO INVESTIGATE THE EXPRESSION LEVEL OF GRAF GENE IN THOSE PATIENTS WITH MYELOID MALIGNANCIES INCLUDING ACUTE MYELOID LEUKEMIA (AML), MYELODYSPLASTIC SYNDROME (MDS) AND CHRONIC MYELOID LEUKEMIA (CML). METHODS: THE EXPRESSION LEVELS OF GRAF TRANSCRIPT WERE DETERMINED IN 94 PATIENTS USING REAL-TIME QUANTITATIVE PCR (RQ-PCR). CLINICAL AND LABORATORY DATA OF THESE PATIENTS WERE COLLECTED AND ANALYZED. RESULTS: THE SIGNIFICANTLY DECREASED LEVEL OF GRAF TRANSCRIPT WAS OBSERVED IN THREE MYELOID MALIGNANCIES COMPARED TO CONTROLS. WITHIN AML, THERE WAS NO DIFFERENCE IN THE LEVEL OF GRAF TRANSCRIPT AMONG DIFFERENT FAB SUBTYPES (P > 0.05). DIFFERENCE WAS NOT OBSERVED IN THE AMOUNT OF GRAF MRNA BETWEEN CML AT CHRONIC PHASE AND CONTROLS. AS CML PROGRESSED, GRAF TRANSCRIPT SIGNIFICANTLY DECREASED. IN MDS, THREE CASES WITH 5Q DELETION HAD LOWER GRAF TRANSCRIPT THAN FOUR WITHOUT 5Q DELETION (MEDIAN 0.76 VS 2.99) (P > 0.05). CONCLUSION: OUR RESULTS DEMONSTRATE THAT THE GRAF TRANSCRIPT IS DECREASED IN MYELOID MALIGNANCIES.	2010	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
18 2327  32 EPIGENETIC REGULATION OF HUMAN CANCER/TESTIS ANTIGEN GENE, HAGE, IN CHRONIC MYELOID LEUKEMIA. BACKGROUND AND OBJECTIVES: CANCER TESTIS ANTIGENS (CTA) PROVIDE ATTRACTIVE TARGETS FOR CANCER-SPECIFIC IMMUNOTHERAPY. ALTHOUGH CTA GENES ARE EXPRESSED IN SOME NORMAL TISSUES, SUCH AS THE TESTIS, THIS IMMUNOLOGICALLY PROTECTED SITE LACKS MHC I EXPRESSION AND AS SUCH, DOES NOT PRESENT SELF ANTIGENS TO T CELLS. TO DATE, CTA GENES HAVE BEEN SHOWN TO BE EXPRESSED IN A RANGE OF SOLID TUMORS VIA DEMETHYLATION OF THEIR PROMOTER CPG ISLANDS, BUT RARELY IN CHRONIC MYELOID LEUKEMIA (CML) OR OTHER HEMATOLOGIC MALIGNANCIES. DESIGN AND METHODS: IN THIS STUDY, THE METHYLATION STATUS OF THE HAGE CTA GENE PROMOTER WAS ANALYZED BY QUANTITATIVE METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MSP) AND SEQUENCING IN FOUR PHILADELPHIA-POSITIVE CELL LINES (TCC-S, K562, KU812 AND KYO-1) AND IN CML SAMPLES TAKEN FROM PATIENTS IN CHRONIC PHASE (CP N=215) OR BLAST CRISIS (BC N=47). HAGE EXPRESSION WAS ASSESSED BY QUANTITATIVE REVERSE TRANSCRIPTASE-POLYMERASE CHAIN REACTION. RESULTS: THE TCC-S CELL LINE SHOWED DEMETHYLATION OF HAGE THAT WAS ASSOCIATED WITH OVER-EXPRESSION OF THIS GENE. HAGE HYPOMETHYLATION WAS SIGNIFICANTLY MORE FREQUENT IN BC (46%) THAN IN CP (22%) (P=0.01) AND WAS CORRELATED WITH HIGH EXPRESSION LEVELS OF HAGE TRANSCRIPTS (P<0.0001). OF NOTE, IN CP-CML, EXTENSIVE HAGE HYPOMETHYLATION WAS ASSOCIATED WITH POORER PROGNOSIS IN TERMS OF CYTOGENETIC RESPONSE TO INTERFERON (P=0.01) OR IMATINIB (P=0.01), MOLECULAR RESPONSE TO IMATINIB (P=0.003) AND PROGRESSION-FREE SURVIVAL (P=0.05). INTERPRETATIONS AND CONCLUSION: THE METHYLATION STATUS OF THE HAGE PROMOTER DIRECTLY CORRELATES WITH ITS EXPRESSION IN BOTH CML CELL LINES AND PATIENTS AND IS ASSOCIATED WITH ADVANCED DISEASE AND POOR OUTCOME.	2007	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
19 4907  26 P53 ABERRATIONS DO NOT PREDICT INDIVIDUAL RESPONSE TO FLUDARABINE IN PATIENTS WITH B-CELL CHRONIC LYMPHOCYTIC LEUKAEMIA IN ADVANCED STAGES RAI III/IV. ABNORMALITIES OF P53 HAVE BEEN ASSOCIATED WITH SHORT SURVIVAL AND NON-RESPONSE TO THERAPY IN CHRONIC LYMPHOCYTIC LEUKAEMIA (CLL). WE HAVE EVALUATED THE RATE OF RESPONSE TO FLUDARABINE AS FIRST-LINE THERAPY IN 54 PATIENTS WITH ADVANCED STAGE CLL, ANALYSING THE CYTOGENETIC PROFILE, ABERRATIONS IN P53, INCLUDING THE METHYLATION STATUS OF ITS PROMOTER, AND THE IMMUNOGLOBULIN HEAVY-CHAIN VARIABLE-REGION (IGVH) MUTATION STATUS. ACCORDING TO THE ADVANCED STAGE OF THE DISEASE IN THIS SERIES, 75% OF PATIENTS PRESENTED GENETIC ABERRATIONS ASSOCIATED WITH POOR PROGNOSIS: DEL(17P) AND/OR DEL(11Q), AND NO-MUTATED IGVH GENES. TEN PATIENTS (18.5%) HAD METHYLATION IN THE PROMOTER REGION OF P53. EIGHTY-THREE PER CENT OF PATIENTS TREATED ACHIEVED A RESPONSE, WITH A HIGH RATE OF COMPLETE REMISSION (47.6%). ALTHOUGH WE FOUND A SIGNIFICANT CORRELATION BETWEEN FAILURES AND THE PRESENCE OF P53 ABERRATIONS (P = 0.0065), EITHER WITH METHYLATION (P = 0.018) OR DELETION (P = 0.015), 64% OF THE PATIENTS WITH ABERRATIONS IN THIS GENE RESPONDED TO TREATMENT (11/17), SUGGESTING THAT FLUDARABINE INDUCES HIGH REMISSION RATES, EVEN IN THESE PATIENTS. THIS IS THE FIRST TIME THAT THE SIGNIFICANCE OF P53 PROMOTER METHYLATION STATUS IS DESCRIBED IN THIS PATHOLOGY, AND OUR DATA SUPPORT THAT THIS EPIGENETIC PHENOMENON COULD BE INVOLVED IN THE PATHOGENESIS AND CLINICAL EVOLUTION OF CLL.	2005	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
20 4549  30 MUTATION ANALYSIS OF THERAPY-RELATED MYELOID NEOPLASMS. WE ANALYZED THE GENETIC MUTATION STATUS OF 13 PATIENTS WITH THERAPY-RELATED MYELOID NEOPLASMS (T-MN). CONSISTENT WITH PREVIOUS REPORTS, T-MN CELLS PREFERENTIALLY ACQUIRED MUTATIONS IN TP53 AND EPIGENETIC MODIFYING GENES, INSTEAD OF MUTATIONS IN TYROSINE KINASE AND SPLICEOSOME GENES. FURTHERMORE, WE COMPARED THE MUTATION STATUS OF THREE T-MN CELLS WITH EACH OF THE INITIAL LYMPHOID MALIGNANT CELLS, AND IDENTIFIED COMMON MUTATIONS AMONG T-MN AND THE INITIAL MALIGNANT CELLS IN TWO PATIENTS. IN A PATIENT WHO DEVELOPED CHRONIC MYELOMONOCYTIC LEUKEMIA (CMML) AFTER FOLLICULAR LYMPHOMA (FL), TET2 MUTATION WAS IDENTIFIED IN BOTH CMML AND FL CELLS. NOTABLY, THE TET2 MUTATION WAS ALSO IDENTIFIED IN PERIPHERAL BLOOD CELLS IN THE DISEASE-FREE PERIOD WITH THE SAME ALLELIC FREQUENCY AS CMML AND FL CELLS, BUT NOT IN A GERM-LINE CONTROL, INDICATING THAT THE TET2 MUTATION OCCURRED SOMATICALLY IN THE INITIATING CLONE FOR BOTH MALIGNANT CELLS. ON THE OTHER HAND, A GERM-LINE MYB MUTATION WAS IDENTIFIED IN A PATIENT WHO DEVELOPED MYELODYSPLASTIC SYNDROMES (MDS) AFTER FL. THESE RESULTS SUGGEST THAT GERM-LINE DEPOSITION AND CLONAL HEMATOPOIESIS ARE CLOSELY ASSOCIATED WITH T-MN SUSCEPTIBILITY; HOWEVER, FURTHER ANALYSIS IS NECESSARY TO CLARIFY THE MECHANISM REQUIRED TO PROVIDE THE INITIATING CLONE WITH LINEAGE COMMITMENT AND CLONAL EXPANSION.	2018