A Yeast Artificial Chromosome Contig That Spans the RB1-D13S31 Interval on Human Chromosome 13 and Encompasses the Frequently Deleted Region in B-cell Chronic Lymphocytic Leukemia

Abstract Loss of the long arm of chromosomes 11 and 13 are the most consistent cytogenetic abnormalities for patients with B-cell chronic lymphocytic leukemia (B-CLL). They suggest the presence of as yet unidentified tumor suppressor genes within well defined minimal deleted regions (MDRs). The use of small vertebrate organisms, such as the zebrafish, as models of diseases associated with chromosomal deletions enables the functional analysis of potential causative genes. Hemopoiesis is well conserved between the zebrafish and human and conserved synteny exists between the two genomes. In this study, the evolutionary conservation between the zebrafish and human genome is investigated for the 13q14 and 11q22-23 regions deleted in B-CLL. Zebrafish orthologs have been identified and radiation hybrid (RH) mapping performed to confirm their chromosomal location and define regions of conserved synteny.We have identified 38 zebrafish orthologs of the human genes in the MDRs in zebrafish cDNA and the syntenic regions for the human deletions in the zebrafish genome. The 13q14 region was syntenic with two main regions in the zebrafish genome, namely linkage group 1 (LG1) and LG9. The majority of zebrafish orthologs to 11q22-23 were found on LG5, LG15 and LG21. One syntenic region, LG9, in the zebrafish genome is of potential interest. Analysis of the smallest critical region of deletion in B-CLL for both 11q22-23 and 13q14 reveals that the human gene equivalents are contained within an area of 22.02 cR on LG9 (approximately 3260 kb). Within LG9, orthologs to two genes to human chromosome 11, three to human chromosome 13 and two chromosome 13 microRNAs (mir-15a and mir-16-1) were identified. The critical region on zebrafish LG9 maps to the MDR for both human chromosomes, suggesting a common ancestry for the B-CLL tumor suppressor genes. This is further supported by analysis of the chicken genome where the same 5 genes from 13q14 and 11q22-23 (C13orf1, RFP2, FLJ11712, FDX1, ARHGAP20) lie within a 10.04 Mb region on chromosome 1. In addition, TILLING for knock-outs of genes in this region of zebrafish embryos will allow analysis of their in vivo potential for lymphoproliferation and may define prime causative genes for B-CLL within human chromosomes 11q and 13q by reverse genetics. Our study provides an explanation for involvement of both 11q and 13q in B-CLL and the potential to develop animal models for this common lymphoproliferative disorder.

Download Full-text

A cosmid and cDNA fine physical map of a human chromosome 13q14 region frequently lost in B-cell chronic lymphocytic leukemia and identification of a new putative tumor suppressor gene, Leu5

FEBS Letters ◽

10.1016/s0014-5793(98)00357-3 ◽

1998 ◽

Vol 426 (2) ◽

pp. 266-270 ◽

Cited By ~ 46

Author(s):

Bagrat Kapanadze ◽

Vladimir Kashuba ◽

Anna Baranova ◽

Omid Rasool ◽

Wim van Everdink ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Human Chromosome ◽

B Cell ◽

Physical Map ◽

Suppressor Gene ◽

Lymphocytic Leukemia ◽

Putative Tumor Suppressor Gene ◽

Putative Tumor Suppressor ◽

Chromosome 13Q14 ◽

Cell Chronic Lymphocytic Leukemia

Download Full-text

Detailed Molecular Delineation of 13q14.3 Loss in B-Cell Chronic Lymphocytic Leukemia

Blood ◽

10.1182/blood.v91.4.1382 ◽

1998 ◽

Vol 91 (4) ◽

pp. 1382-1390 ◽

Cited By ~ 73

Author(s):

Martin M. Corcoran ◽

Omid Rasool ◽

Yie Liu ◽

Arati Iyengar ◽

Dan Grander ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

B Cell ◽

Cpg Islands ◽

Artificial Chromosome ◽

Lymphocytic Leukemia ◽

Deletion Mapping ◽

Polymorphic Dinucleotide ◽

Genomic Probes ◽

Homozygous Deletions ◽

Cell Chronic Lymphocytic Leukemia

Abstract A region of chromosome 13q14.3, telomeric to the Retinoblastoma gene RB-1 is frequently deleted in patients with B-cell chronic lymphocytic leukemia (B-CLL). A cosmid and P1-derived artificial chromosome (PAC) contig spanning over 600 kb has been constructed, which encompasses this locus. The contig clones have been used to order a number of markers along the minimally deleted region and to localize a series of CpG islands corresponding to possible candidate genes. A novel polymorphic dinucleotide repeat, 6E3.2, present in one of the ordered cosmid clones has been isolated for use in deletion mapping studies of patient DNA. Leukemic samples from 229 CLL patients have been screened for loss of heterozygosity using microsatellite markers and analyzed for hemizygous and homozygous deletions by Southern blot techniques using genomic probes selected from cosmids across the region. Hemizygous deletions were found in 31% of cases with an additional 10% showing homozygous loss. The use of these probes has defined the commonly deleted area to less than 130 kb, centromeric to the locus D13S272.

Download Full-text

Minimal region of loss at 13q14 in B-cell chronic lymphocytic leukemia

Blood ◽

10.1182/blood.v88.8.3109.bloodjournal8883109 ◽

1996 ◽

Vol 88 (8) ◽

pp. 3109-3115 ◽

Cited By ~ 71

Author(s):

F Bullrich ◽

ML Veronese ◽

S Kitada ◽

J Jurlander ◽

MA Caligiuri ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Tumor Suppressor ◽

B Cell ◽

Tumor Suppressor Genes ◽

Lymphocytic Leukemia ◽

Allelic Loss ◽

Loss Of Function ◽

Suppressor Genes ◽

Chromosome 13 ◽

Cell Chronic Lymphocytic Leukemia

Allelic loss at nonrandom chromosomal sites is thought to mark the position of tumor suppressor genes involved in the pathogenesis and progression of human malignancies. Solid tumors in particular have been found to harbor multiple genetic changes resulting in loss of function mutations. Tumor suppressor genes have also been found to be involved in the progression of lymphoid tumors. Previous reports have suggested the involvement of a tumor suppressor gene located on the long arm of chromosome 13, between the retinoblastoma (RB) and D13S25 loci, in the pathogenesis and or progression of more than 40% of B-cell chronic lymphocytic leukemia (B-CLL), a common lymphoid malignancy whose molecular etiology remains largely unknown. In the present study, we report the construction and characterization of a YAC contig spanning a region of approximately 3 cM between the RB gene and the D13S31 locus. We also screened 60 paired normal/tumor B-CLL samples for allelic loss on chromosome 13 with nine microsatellite markers located between RB and D13S25. This analysis has allowed us to narrow the smallest region of loss to a segment of 550 kb located between the 206XF12 and D13S25 markers.

Download Full-text

Construction of a bacterial artificial chromosome (BAC) contig across the minimally deleted region in 13q14.3 in B-cell chronic lymphocytic leukemia

Annals of Human Genetics ◽

10.1046/j.1469-1809.1998.6250401.x ◽

1998 ◽

Vol 62 (5) ◽

pp. 401-409 ◽

Cited By ~ 1

Author(s):

L. A. HAWTHORN ◽

J. K. COWELL

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Bacterial Artificial Chromosome ◽

B Cell ◽

Artificial Chromosome ◽

Lymphocytic Leukemia ◽

Bac Contig ◽

Cell Chronic Lymphocytic Leukemia

Download Full-text

Chromosome 13… new marker for B-cell chronic lymphocytic leukemia

Hereditas ◽

10.1111/j.1601-5223.1988.tb00684.x ◽

2008 ◽

Vol 108 (1) ◽

pp. 77-84 ◽

Cited By ~ 20

Author(s):

LORE ZECH ◽

HÅKAN MELLSTEDT

Keyword(s):

Chronic Lymphocytic Leukemia ◽

B Cell ◽

Lymphocytic Leukemia ◽

Chromosome 13 ◽

Cell Chronic Lymphocytic Leukemia

Download Full-text

11q Deletions Identify a New Subset of B-Cell Chronic Lymphocytic Leukemia Characterized by Extensive Nodal Involvement and Inferior Prognosis

Blood ◽

10.1182/blood.v89.7.2516 ◽

1997 ◽

Vol 89 (7) ◽

pp. 2516-2522 ◽

Cited By ~ 276

Author(s):

Hartmut Döhner ◽

Stephan Stilgenbauer ◽

Michael R. James ◽

Axel Benner ◽

Traudel Weilguni ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

B Cell ◽

Mononuclear Cells ◽

Yeast Artificial Chromosome ◽

Lymphoproliferative Disorders ◽

Lymphocytic Leukemia ◽

Mediastinal Lymphadenopathy ◽

Chromosome 11 ◽

Significant Difference ◽

Cell Chronic Lymphocytic Leukemia

Abstract Deletions of the long arm of chromosome 11 (11q) are one of the most frequent structural chromosome aberrations in various types of lymphoproliferative disorders. However, in most conventional chromosome banding studies of B-cell chronic lymphocytic leukemia (B-CLL), 11q deletions were not identified as a frequent aberration. The objective of this study was to analyze the frequency and clinical impact of 11q deletions in B-CLL by interphase cytogenetics using fluorescence in situ hybridization (FISH). Mononuclear cells from 214 patients with B-CLL were studied by FISH using the yeast artificial chromosome (YAC) clone 755b11 from chromosome region 11q22.3-q23.1; we previously showed that this clone was contained within a 2- to 3-Mb sized segment of 11q commonly deleted in lymphoproliferative disorders. Forty-three of the 214 (20%) tumors exhibited 11q deletions; 11q deletions were the second most frequent chromosome aberration following 13q14 (RB1 and/or D13S25) deletions (45%); they were more frequent than trisomy 12 (15%) or deletion of 17p (TP53 gene) (10%). Patients with 11q deletions were younger (P = .01) and had more advanced clinical stages (P = .01). 11q deletions were associated with extensive peripheral, abdominal, and mediastinal lymphadenopathy (P < .001). Patients with 11q deletions had a more rapid disease progression as shown by a shorter treatment-free interval (9 months v 43 months; P < .001). The prognostic effect of 11q deletion on survival strongly depended on the age: in patients less than 55 years old, the median survival time was significantly shorter in the deletion group (64 months v 209 months; P < .001), whereas in patients ≥ 55 years old there was no significant difference (94 months v 111 months; P = .82). 11q deletions identify a new clinical subset of B-CLL characterized by extensive lymph node involvement. In younger B-CLL patients, this aberration is an important predictor of survival.

Download Full-text