Illumina NovaSeq 6000 paired end sequencing; Single Stem Cell Lineage Analysis of the Leukemic Clones Responsible for Acute Myeloid Leukemia Relapse Based on the Functional and Genomic Properties at Diagnosis
Single Stem Cell Lineage Analysis of the Leukemic Clones Responsible for Acute Myeloid Leukemia Relapse Based on the Functional and Genomic Properties at Diagnosis
peripheral blood (PB) and Bone Marrow (BM) obtained from the Rambam Bio-Bank, from AML patients from both diagnosis (Dx) and Rx were studied.  Initially CD34+ enriched fraction was sorted into 2 populations of CD33+ and CD33- reflecting leukemic and normal progenitors, respectively. This was followed by sorting into different progenitor populations to yield homogenous populations from which subclones will be generated. This includes hematopoietic stem cell-multi potent progenitor (HSC/MPP), common myeloid progenitor (CMP), granulocyte myeloid progenitor (GMP) and multipotent/common lymphoid progenitor (MLP/CLP). each subpopulation was further sorted into single cell or alternatively undergone single cell subclone formation. Both single cells and single cell derived subclones were further sequenced. Samples that were grown in mouse are xenografts as interfemoral transplantation of hematopoietic stem and progenitor cells into Nod-scid immune suppressed female mice MDA based single cell WGA kit as described per sample in Biezuner et al. Genome Research. 2016. Targeted enrichment using duplex Molecular Inversion Probes protocol as described in (https://doi.org/10.1016/j.crmeth.2021.100054)