Leukemia

About this book

Recent advances in molecular and cellular biology techniques have significantly improved our ability to detect, monitor, model and study the underlying molecular basis and pathogenesis of leukemia, yet we are still in an early discovery stage and much more work is needed in order to develop better strategies to diagnose, classify and treat this biologically and clinically diverse disease. In Leukemia: Methods and Protocols, expert researchers bring together a wide range of state-of-the-art laboratory methods and detailed protocols that are useful for both clinical and basic research scientists working on the disease. The volume provides techniques for prenatal backtracking of leukemic clone, molecular diagnosis, detection of genome-wide genetic abnormalities and profiling, identification of unknown fusion genes, monitoring of minimal residual diseases, disease modeling using murine and human primary hematopoietic cells, studying of normal and malignant hematopoiesis, identification of interacting partners with leukemia associated oncoproteins, and global characterization of genome-wide epigenetic changes in leukemic cells. Written in the highly successful Methods in Molecular Biology™ series format, the convenient chapters contain brief introductions, lists of the necessary materials, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.

Comprehensive and cutting-edge, Leukemia: Methods and Protocols will help researchers to advance knowledge and have a better understanding of the disease, which will ultimately facilitate development of anti-cancer therapy and improve quality of life for patients.

Written for: Molecular and cellular biologists, hematologists, cancer researchers

Keywords:

• Anti-cancer therapy
• Disease modeling
• Experimental hematology
• Hematological malignancies
• Molecular diagnosis

Table of contents

1 An Overview: From Discovery of Candidate Mutations to Disease Modelling and Transformation Mechanisms of Acute Leukemia
Chi Wai Eric So

2 Backtracking of Leukemic Clones to Birth
Joseph Wiemels, Michelle Kang, and Mel Greaves

3 Fluorescence in situ Hybridization (FISH) as a Tool for the Detection of Significant Chromosomal Abnormalities in Childhood Leukaemia
Zoë J. Konn, Sarah L. Wright, Kerry E. Barber, and Christine J. Harrison

4 Specialized Fluorescence in situ Hybridization (FISH) Techniques for Leukemia Research
Lyndal Kearney and Sue Colman

5 LDI-PCR: Identification of Known and Unknown Gene Fusions of the Human MLL Gene
Claus Meyer and Rolf Marschalek

6 Panhandle PCR Approaches to Cloning MLL Genomic Breakpoint Junctions and Fusion Transcript Sequences
Blaine W. Robinson and Carolyn A. Felix

7 MRD Detection in Acute Lymphoblastic Leukemia Patients Using Ig/TCR Gene Rearrangements as Targets for Real-Time Quantitative PCR
Vincent H.J. van der Velden and Jacques J.M. van Dongen

8 Array-Based Comparative Genomic Hybridization as a Tool for Analyzing the Leukemia Genome
Jon C. Strefford and Helen Parker

9 Application of SNP Genotype Arrays to Determine Somatic Changes in Cancer
Manu Gupta and Bryan D. Young

10 Retroviral/Lentiviral Transduction and Transformation Assay
Bernd B. Zeisig and Chi Wai Eric So

11 Gene Expression Profiling of Leukemia Stem Cells
Andrei V. Krivtsov, Yingzi Wang, Zhaohui Feng, and Scott A. Armstrong

12 Humanized Model to Study Leukemic Stem Cells
Dominique Bonnet

13 Model Systems for Examining Effects of Leukemia Associated Oncogenes in Primary Human CD34+ Cells via Retroviral Transduction
Mark Wunderlich and James C. Mulloy

14 Ex-vivo Assays to Study Self-Renewal and Long-Term Expansion of Genetically Modified Primary Human Acute Myeloid Leukemia Stem Cells
Jan Jacob Schuringa and Hein Schepers

15 Identification and Characterization of Hematopoietic Stem and Progenitor Cell Populations in Mouse Bone Marrow by Flow Cytometry
Jenny Yeung and Chi Wai Eric So

16 In vitro Differentiation of ES Cells as a Model of Early Hematopoietic Development
Patrycja Sroczynska, Christophe Lancrin, Stella Pearson, Valerie Kouskoff, and Georges Lacaud

17 Analysis of the Mouse Placenta as a Hematopoietic Stem Cell Niche
Katrin Ottersbach and Elaine Dzierzak

18 Identification of Protein Interaction Partners by the Yeast Two Hybrid System
Maria-Paz Garcia-Cuellar, Deniz Mederer, and Robert K. Slany

19 Complete Array of HOX Gene Expression by RQ-PCR
Glenda J. McGonigle, Terence R. Lappin, and Alexander Thompson

20 Genome-Wide Determination of DNA Methylation by HpaII Tiny Fragment Enrichment by Ligation-Mediated PCR (HELP) for the Study of Acute Leukemias
Maria E. Figueroa, Ari M. Melnick, and John M. Greally

21 Chromatin Immunoprecipitation (ChIP) for Analysis of Histone Modifications and Chromatin Associated Proteins
Thomas A. Milne, Keji Zhao, and Jay L. Hess