Genetic Susceptibility to Therapy-Related Acute Murine Promyelocytic Leukemia.

Abstract Therapy-related acute myelogenous leukemia (t-AML) is an important late adverse effect of alkylator chemotherapy. Susceptibility to t-AML has a genetic component, yet the specific genes and genetic variations that influence susceptibility are poorly understood. Our lab previously identified mouse strains that are susceptible (SWR/J) or resistant (C57BL/6J and C3H/HeJ) to t-AML induced by the alkylator, ethyl-N-nitrosourea (ENU). To study the genetic basis of these differences, we performed an F2 intercross between susceptible and resistant strains. A single copy of the hCG-PML-RARa (PR) transgene was bred into each mouse. PR is an initiating factor for acute promyelocytic leukemia, but requires cooperating mutations for full leukemic transformation. This provides a platform to detect gene X transgene (PR) and gene X environment (ENU) interactions that promote leukemogenesis. F2 mice were treated (n=141) or untreated (n=141) with ENU and sacrificed and analyzed when moribund. We also analyzed untreated PR+ (n=24) mice from the resistant C57BL/6J X C3H/HeJ (B6C3F1) background. Untreated B6C3F1 PR+ mice developed lethal myeloid leukemia (characterized by splenomegaly > 0.25g, WBC > 30 K/uL, and increased immature myeloid precursors in PB, BM, and spleen) with an incidence of 12.5% and a latency of 234 days. By contrast, 79.4% of untreated PR+ F2 mice developed myeloid leukemia with a latency of 108 days and median survival of 238 days. The earlier onset and increased incidence of leukemia in F2 mice confirm that SWR/J alleles confer increased susceptibility to AML. ENU treatment further increased the leukemia incidence (90.4% vs. 79.4%, p<0.0001 by logrank) and shortened the median survival (168 vs. 238 days) of PR+ F2 mice. F2 mice were genotyped using 357 informative SNPs across the genome to facilitate quantitative trait locus (QTL) mapping. QTL analysis was performed using leukemia-free survival, spleen weight, and WBC as quantitative traits. Because extramedullary hematopoeisis and increased WBC are markers of poor prognosis in human AML, we reasoned that identification of modifier alleles for these traits might also have potential clinical relevance. QTL analysis revealed five peaks associated with survival, 5 with spleen weight, and 3 with WBC. The 1 LPR (likelihood probability ratio) confidence intervals for the QTL range in size from 10 to 50 Mbp. Each region contains between 100 and 750 genes. The most significant peak (LPR=3.94) is a survival QTL on chromosome 1 from 93.4 to 120.5 Mbp that retains significance at the genome wide level. The QTL effect is large in ENU-treated animals but not discernible in untreated animals. Both the additive effect (−0.51, SE=0.17) and the dominant effect (0.62, SE=0.20) are significant at p<0.05. Genes with potential connections to leukemogenesis within this region include a serpin cluster, genes involved in apoptosis (Bcl2, Bok, Pdcd1 Phlpp), and cell cycle genes (Sept2 and Clasp1). Ongoing studies are focused on candidate gene evaluation and fine-mapping the QTL regions to identify the QTL genes and their variants. Validation of these genes in therapy-related leukemogenesis should provide a better understanding of t-AML susceptibility and lead to strategies that moderate t-AML risk in susceptible individuals.

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Quantitative trait loci associated with susceptibility to therapy-related acute murine promyelocytic leukemia in hCG-PML/RARA transgenic mice

Blood ◽

10.1182/blood-2008-01-132084 ◽

2008 ◽

Vol 112 (4) ◽

pp. 1434-1442 ◽

Cited By ~ 10

Author(s):

Ryan K. Funk ◽

Taylor J. Maxwell ◽

Masayo Izumi ◽

Deepa Edwin ◽

Friederike Kreisel ◽

...

Keyword(s):

Quantitative Trait ◽

Complex Trait ◽

Myelogenous Leukemia ◽

Mouse Strains ◽

Spleen Weight ◽

Nucleotide Polymorphisms ◽

Treatment Regimens ◽

Multiple Loci ◽

Acute Myelogenous ◽

Trait Locus

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Establishment of baseline toxicity expectations with standard frontline chemotherapy in acute myelogenous leukemia

Blood ◽

10.1182/blood-2007-06-095844 ◽

2007 ◽

Vol 110 (10) ◽

pp. 3547-3551 ◽

Cited By ~ 62

Author(s):

Ehab Atallah ◽

Jorge Cortes ◽

Susan O'Brien ◽

Sherry Pierce ◽

Mary Beth Rios ◽

...

Keyword(s):

Adverse Events ◽

Myeloid Leukemia ◽

Febrile Episode ◽

Promyelocytic Leukemia ◽

Myelogenous Leukemia ◽

Control Group ◽

Intensive Chemotherapy ◽

Serious Adverse Events ◽

Acute Myelogenous ◽

Grade 3

Abstract The rates of expected serious adverse events in patients with acute leukemia on chemotherapy far exceed those in patients with solid tumors. Regulatory authorities require similar reporting criteria, which overburden the investigators and infrastructure with unnecessary documentation. To establish a baseline for expected toxicities before and during leukemia therapy, we reviewed 1534 adults with acute myeloid leukemia (AML; excluding acute promyelocytic leukemia) from 1990 to 2006 who received frontline intensive chemotherapy; 723 (47%) were 60 years or older. Prior to therapy, grade 3/4 cytopenias were observed in 86% of patients. All patients developed one or more grade 3/4 cytopenias during therapy, and more than 90% had a febrile episode. Admission to the intensive care unit, mechanical ventilation, and dialysis were required in 28%, 16%, and 7%, respectively. Mortality during induction, 2-week mortality, and 6-week mortality were 20%, 5%, and 16%, respectively. Grade 3/4 renal or hepatic toxicities were observed in 3% and 22% of patients, respectively. Other grade 3 or 4 toxicities were also common before treatment and during therapy. This paper establishes a baseline toxicity rate for patients with AML during induction therapy, and this could be used as a control group for future reference. Guidelines for reporting adverse events in leukemia studies should be revisited.

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Decitabine-Based Salvage Therapy in Adults with Acute Myeloid Leukemia.

Blood ◽

10.1182/blood.v114.22.2063.2063 ◽

2009 ◽

Vol 114 (22) ◽

pp. 2063-2063 ◽

Cited By ~ 1

Author(s):

Ellen K Ritchie ◽

Jon Arnason ◽

Eric J. Feldman ◽

Usama S Gergis ◽

Sebastian A Mayer ◽

...

Keyword(s):

Bone Marrow ◽

Acute Myeloid Leukemia ◽

Median Survival ◽

Salvage Therapy ◽

Myeloid Leukemia ◽

Gemtuzumab Ozogamicin ◽

Myelogenous Leukemia ◽

Rank Test ◽

Off Label ◽

Acute Myeloid

Abstract Abstract 2063 Poster Board II-40 Introduction: Adults with primary refractory or relapsed acute myeloid leukemia (AML) have a poor prognosis with complete responses to salvage therapy from 13-15% and a median survival of 1.5-6 months[1, 2]. From 2006-2009, 79 patients with relapsed or refractory acute myeloid leukemia were given salvage chemotherapy with decitabine 20mg/m2 daily for 10 days or decitabine 20mg/m2 × 5 days with gemtuzumab ozogamicin (GO) 3mg/m2 on day 5 at Weill Cornell Medical Center. Methods: Medical records of 79 patients who received decitabine-based salvage therapy were reviewed from September 2006 through July 2009 at Weill Cornell Medical College. Survival was calculated by the Kaplan Meir method and differences in survival calculated by the log-rank test using STATA software. Results: Twenty-five patients received decitabine-based therapy as first salvage, 32 patients as second salvage, and 22 patients as third or greater salvage. Fifty-one patients were treated with decitabine/GO and 29 patients received decitabine alone. Median age of patients was 65.5 years with a range of 24 -89 years (first salvage 75 years, second salvage 62 years, and third or greater salvage 64 years). Median survival of all patients was 205 days, range 7-732 days. Overall 34% patients responded: 16% CR (<5% blasts in bone marrow, recovery ANC >1000 and Plts > 100,000) with median survival not yet reached; 5% CRp (< 5% blasts in bone marrow, ANC >1000 and plts < 100,000) with median survival 223 days; 13% PR (blasts 6-11%) with median survival 205 days; and 66% no response with median survival 118 days. Patients receiving first salvage had median survival 181 days with CR 13%, CRp 6%, PR 17%. Patients receiving second salvage had median survival 207 days with CR 9%, CRp 4%, PR 9%. Patients with third or greater salvage had median survival 209 days with CR 23%, CRp 0%, PR 14%. Patients receiving decitabine alone had a median survival of 209 days and those receiving decitabine/GO had a median survival of 177 days but the difference was not significant. Median survival for normal, favorable, intermediate and unfavorable cytogenetics was 282, 224, 157 and 176 days respectively (p=0.06). Conclusions: Decitabine-based treatment for relapsed and refractory AML is a low intensity alternative that has activity rivaling more intensive regimens. This retrospective study suggests that further investigation of decitabine-based salvage is warranted. 1. Sievers, E.L., et al., Efficacy and safety of gemtuzumab ozogamicin in patients with CD33-positive acute myeloid leukemia in first relapse. J Clin Oncol, 2001. 19(13): p. 3244-54. 2. Giles, F., et al., Outcome of patients with acute myelogenous leukemia after second salvage therapy. Cancer, 2005. 104(3): p. 547-54. Disclosures: Off Label Use: Phase I trial of decitabine in AML is off-label .

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Whole Arm Duplication of 1q in Myeloid Neoplasm, with Emphasis On Derivative (1;7)(q10;p10).

Blood ◽

10.1182/blood.v114.22.4238.4238 ◽

2009 ◽

Vol 114 (22) ◽

pp. 4238-4238

Author(s):

Patricia T. Greipp ◽

Curtis A Hanson ◽

Ruchi G. Sharma ◽

Lai P. Nguyen ◽

Ryan A Knudson ◽

...

Keyword(s):

Bone Marrow ◽

Acute Myeloid Leukemia ◽

Median Survival ◽

Mayo Clinic ◽

Myeloid Leukemia ◽

Cytogenetic Study ◽

Chromosome 7 ◽

Chromosome 1 ◽

Hematologic Neoplasm ◽

Acute Myeloid

Abstract Abstract 4238 Background Duplication of the long arm of chromosome 1 (1q) is a recurrent finding in hematologic neoplasms. It is most commonly observed as an unbalanced whole arm translocation with chromosome 7, resulting in concurrent deletion of 7q (ISCN as der(1;7)(q10q10)). It is less frequently observed as an unbalanced translocation with a chromosome other than chromosome 7 or as a duplication of only a segment of 1q. The der(1;7) has been described in myelodysplastic syndrome (MDS), and acute myeloid leukemia (AML), and is reportedly less commonly associated with myeloproliferative neoplasms (MPN). The effect of this rearrangement on prognosis is unclear, although it is classified by the International Scoring System (IPSS) as a karyotypic variant of del(7q)/-7 and is thus assigned poor risk. We have initiated a thorough review of all cases of duplication of the long arm of chromosome 1 seen at our institution. Methods After approval from the Institutional Review Board, all cases seen at Mayo Clinic over a 20 year period (1989-2009) were searched for a duplication of the long arm of chromosome 1. The sole exclusion criterion was a complex karyotype (3 or more independent chromosome abnormalities in the clone). Cytogenetic and hematopathology review was done and relevant clinical data were abstracted from the Mayo Clinic record. Survival of MDS patients and MPN patients was determined from time of cytogenetic diagnosis of der(1;7) until death or last contact with the patient, using the Kaplan-Meier method. Results Among 23,375 unique patients at Mayo Clinic with a cytogenetic study for hematologic neoplasm, 229 (0.98%) patients carried a duplication of 1q. Within this group, 99 (0.42%) had duplication of part of 1q, 123 (0.53%) carried a whole arm duplication of 1q, and 7 patients carried an isochromosome 1q (resulting in 4 copies of 1q). The most common karyotype among those with a whole arm duplication of 1q was der(1;7) (n=84), followed by similar whole arm translocations involving chromosomes 9 and 19 (n=7 each) or chromosomes 12, 13, 14, 15, 16, 17, 18, 20, 21, or 22 (n=1 to 6 each). We focused on 65 der(1;7) patients who had adequate hematopathological material that could be reviewed and sub-classified according to WHO criteria. MDS was identified in 30 patients and MPN in 20. MDS (therapy related, RARS, RAEB-1 or 2) was seen in 9 patients (14%), MDS/ refractory cytopenia with multilineage dysplasia (RCMD) in 19 (29%). MDS/Myeloproliferative neoplasm (MPN) in 2 (3%), MPN not otherwise classified in 5 (8%), and MPN / primary myelofibrosis (including 1 post-polycythemic and one post-thromobocythemic) (PMF) in 15 (23%). The bone marrow was apparently normal in 5 patients (8%), CLL was noted in 3 (5%), and 1 each exhibited features of systemic mast cell disease or myeloma. Acute myeloid leukemia (AML) was identified 5 patients (8%). The M:F ratio of the der(1;7) patients was 2.8:1. The median age at occurrence of the der(1;7) was 63 years (range: 13-87 years). All 5 AML patients have died (median survival 9 months). Of the MDS and MPN patients, the median survival was 2.2 and 2.8 years, respectively, from the time of cytogenetic diagnosis. Overall survival was not significantly different between the MDS and MPN group, and they exhibited 30% survival after 5 years (see Figure). Conclusions Duplication of 1q was observed in nearly 1% of the Mayo Clinic patients who had a bone marrow cytogenetic study for a hematologic neoplasm. Among those with a der(1;7), AML was uncommon but MDS and MPN were similar in frequency. Median survival of 2.2 to 2.8 years for the MDS and MPN group justifies assignment to a more favorable group than the current IPSS assignment. Disclosures: No relevant conflicts of interest to declare.

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Requirement of c-Myb for p210BCR/ABL-dependent transformation of hematopoietic progenitors and leukemogenesis

Blood ◽

10.1182/blood-2007-08-105072 ◽

2008 ◽

Vol 111 (9) ◽

pp. 4771-4779 ◽

Cited By ~ 42

Author(s):

Maria Rosa Lidonnici ◽

Francesca Corradini ◽

Todd Waldron ◽

Timothy P. Bender ◽

Bruno Calabretta

Keyword(s):

Median Survival ◽

Colony Formation ◽

Myeloid Leukemia ◽

Blast Crisis ◽

Cyclin B1 ◽

Myelogenous Leukemia ◽

Hematopoietic Progenitors ◽

Myb Gene ◽

Blast Cells ◽

Marrow Cells

Abstract The c-Myb gene encodes a transcription factor required for proliferation and survival of normal myeloid progenitors and leukemic blast cells. Targeting of c-Myb by antisense oligodeoxynucleotides has suggested that myeloid leukemia blasts (including chronic myelogenous leukemia [CML]–blast crisis cells) rely on c-Myb expression more than normal progenitors, but a genetic approach to assess the requirement of c-Myb by p210BCR/ABL-transformed hematopoietic progenitors has not been taken. We show here that loss of a c-Myb allele had modest effects (20%-28% decrease) on colony formation of nontransduced progenitors, while the effect on p210BCR/ABL-expressing Lin− Sca-1+ and Lin− Sca-1+Kit+ cells was more pronounced (50%-80% decrease). Using a model of CML-blast crisis, mice (n = 14) injected with p210BCR/ABL-transduced p53−/−c-Mybw/w marrow cells developed leukemia rapidly and had a median survival of 26 days, while only 67% of mice (n = 12) injected with p210BCR/ABL-transduced p53−/−c-Mybw/d marrow cells died of leukemia with a median survival of 96 days. p210BCR/ABL-transduced c-Mybw/w and c-Mybw/d marrow progenitors expressed similar levels of the c-Myb–regulated genes c-Myc and cyclin B1, while those of Bcl-2 were reduced. However, ectopic Bcl-2 expression did not enhance colony formation of p210BCR/ABL-transduced c-Mybw/d Lin−Sca-1+Kit+ cells. Together, these studies support the requirement of c-Myb for p210BCR/ABL-dependent leukemogenesis.

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Pseudo-Chediak-Higashi anomaly in a case of acute myeloid leukemia: electron microscopic studies

Blood ◽

10.1182/blood.v54.4.863.863 ◽

1979 ◽

Vol 54 (4) ◽

pp. 863-871 ◽

Cited By ~ 20

Author(s):

M Tulliez ◽

JP Vernant ◽

J Breton-Gorius ◽

M Imbert ◽

C Sultan

Keyword(s):

Electron Microscopy ◽

Lymph Nodes ◽

Acute Promyelocytic Leukemia ◽

Myeloid Leukemia ◽

Promyelocytic Leukemia ◽

Myelogenous Leukemia ◽

Electron Microscopic ◽

Microscopic Studies ◽

Acute Myelogenous ◽

Chediak Higashi Syndrome

Abstract The formation and fine structure of giant granules in neutrophil promyelocytes of a patient with a variant of acute myelogenous leukemia were investigated by electron microscopy. The patient presented with large lymph nodes and disseminated intravascular coagulation (DIC). By light microscopy, numerous giant granules, resembling those of Chediak- Higashi syndrome (CHS), were present, but Auer bodies could not be found. By electron microscopy, these giant granules were seen to be formed by fusion of azurophilic granules, as in CHS; however, they were different from the large granules of CHS, since they contained numerous microcrystalline structures like those of Auer bodies. However, the crystalline cores of these granules exhibited a periodicity different from that of Auer bodies of acute promyelocytic leukemia. This clinical and hematologic syndrome (giant granules, enlarged lymph nodes, and DIC may represent a variant of acute promyelocytic leukemia.

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Pseudo-Chediak-Higashi anomaly in a case of acute myeloid leukemia: electron microscopic studies

Blood ◽

10.1182/blood.v54.4.863.bloodjournal544863 ◽

1979 ◽

Vol 54 (4) ◽

pp. 863-871

Author(s):

M Tulliez ◽

JP Vernant ◽

J Breton-Gorius ◽

M Imbert ◽

C Sultan

Keyword(s):

Electron Microscopy ◽

Lymph Nodes ◽

Acute Promyelocytic Leukemia ◽

Myeloid Leukemia ◽

Promyelocytic Leukemia ◽

Myelogenous Leukemia ◽

Electron Microscopic ◽

Microscopic Studies ◽

Acute Myelogenous ◽

Chediak Higashi Syndrome

The formation and fine structure of giant granules in neutrophil promyelocytes of a patient with a variant of acute myelogenous leukemia were investigated by electron microscopy. The patient presented with large lymph nodes and disseminated intravascular coagulation (DIC). By light microscopy, numerous giant granules, resembling those of Chediak- Higashi syndrome (CHS), were present, but Auer bodies could not be found. By electron microscopy, these giant granules were seen to be formed by fusion of azurophilic granules, as in CHS; however, they were different from the large granules of CHS, since they contained numerous microcrystalline structures like those of Auer bodies. However, the crystalline cores of these granules exhibited a periodicity different from that of Auer bodies of acute promyelocytic leukemia. This clinical and hematologic syndrome (giant granules, enlarged lymph nodes, and DIC may represent a variant of acute promyelocytic leukemia.

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Genetic analysis of blood pressure in C3H/HeJ and SWR/J mice

Physiological Genomics ◽

10.1152/physiolgenomics.00212.2003 ◽

2004 ◽

Vol 17 (2) ◽

pp. 215-220 ◽

Cited By ~ 46

Author(s):

Keith DiPetrillo ◽

Shirng-Wern Tsaih ◽

Susan Sheehan ◽

Conrado Johns ◽

Peter Kelmenson ◽

...

Keyword(s):

Blood Pressure ◽

Confidence Interval ◽

Qtl Analysis ◽

Inbred Strains ◽

Chromosome 1 ◽

Causal Gene ◽

Complex Phenotype ◽

Males And Females ◽

Trait Locus ◽

Genomic Regions

Hypertension is a complex phenotype induced by multiple environmental and genetic factors. Quantitative trait locus (QTL) analysis is a powerful method for identifying genomic regions underlying complex diseases. We conducted a QTL analysis of blood pressure in mice using 217 F2 progeny (males and females) from a cross between the normotensive C3H/HeJ and hypertensive SWR/J inbred strains. Our analysis identified significant QTL controlling blood pressure on chromosome 1 [Chr 1; Bpq8; peak 78 cM; 95% confidence interval 64–106 cM; logarithm of the odds ratio (LOD) 3.5; peak marker D1Mit105] and on Chr 16 ( Bpq9; peak 56 cM; 95% confidence interval 46–58 cM; LOD 3.6; peak marker D16Mit158). Bpq8 was previously identified in a cross between C57BL/6J and A/J mice, and we narrowed this QTL from 42 to 18 cM (95% confidence interval 68–86 cM) by combining the data from these crosses. By examining Bpq8 for regions where ancestral alleles were conserved among the high allele strains (C57BL/6J, SWR/J) and different from the low allele strains (A/J, C3H/HeJ), we identified a 2.3-cM region where the high allele strains shared a common haplotype. Bpq8 is concordant with known QTL in both rat and human, suggesting that the causal gene underlying Bpq8 may be conserved as a disease gene in human hypertension.

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Abstract 1388: Hdlq14 Gene, A New Gene Regulating HDL Levels

Circulation ◽

10.1161/circ.116.suppl_16.ii_285-a ◽

2007 ◽

Vol 116 (suppl_16) ◽

Author(s):

Zhiguang Su ◽

Allison Cox ◽

Yuan Shen ◽

Ioannis Stylianou ◽

Beverly Paigen

Keyword(s):

Qtl Analysis ◽

Complex Traits ◽

Disease Risk ◽

Cardiovascular Disease Risk ◽

Density Lipoprotein ◽

Mouse Strains ◽

F2 Population ◽

New Genes ◽

Trait Locus ◽

Significant Expression

Background . The discovery of new genes responsible for regulation of high-density lipoprotein cholesterol (HDL) has great clinical relevance since increases in HDL can reduce cardiovascular disease risk. Quantitative trait locus (QTL) analysis is a means of finding novel genes that regulate complex traits, such as atherosclerosis and HDL. Hdlq14 and Hdlq15 , two closely linked QTLs for HDL on mouse Chr 1, have been detected by using an intercross between strains C57BL/6 (B6) and 129S1/SvImJ (129). Apoa2 is the gene for Hdlq15 locus, but the gene for Hdlq14 is unknown. Methods: To confirm the Hdlq14 and identify the candidate gene, we performed QTL analysis in a F2 population generated from strains NZB and NZW, which are same at Apoa2 to avoid its strong effect on the nearby QTL. Hdlq14 was further narrowed by several strategies including combining crosses, comparative genomics, and haplotype analysis. The reduced lists of candidate genes were evaluated by their expression or sequence differences between the strains that caused the Hdlq14 . Finally, other HDL crosses, including NZOxNON, B6xC3H, and Pera x D2, were examined to point out the QTL gene. The relationship between the polymorphism at the Hdlq14 gene and HDL was analyzed in 43 genetically diverse mouse strains. Results: Hdlq14 was proved in cross NZBxNZW and the critical interval was reduced from 45 Mb harboring 271 genes to 1.65 Mb containing 15 genes by using bioinformatics tools. Six of these 15 genes have polymorphisms that changed an amino acid; and two genes were found have a significant expression difference between strains B6 and 129. The Hdlq14 gene was further pointed out using HDL QTL identified in crosses including NZOxNON, B6xC3H, and PeraxDBA. In 43 genetically diverse mouse strains, we found that strains with one allele of the Hdlq14 had significantly higher plasma HDL levels than those with the other variant. Conclusions: The Hdlq14 was identified as a new HDL-regulating gene.

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