erization domain or PEST domain or alternatively loss-of-function mutations in FBXW7, a NOTCH1 E3 ubiquitin ligase, increase release or stability of ICN1. This, in turn, leads to transcriptional activation of genes that promote proliferation and survival such as MYC and HES1. Despite a plethora of reports describing mechanisms of NOTCH1 activation in T-ALL, the cell type and context specific role of NOTCH1 activation in the maintenance of therapeutically resistant self-renewing human LIC has not been established. Thus, we sought to examine whether molecularly characterized LIC can be identified among specific hematopoietic subpopulations in pediatric T-ALL without preceding in vitro culture, the role of NOTCH1 activation in LIC propagation, and whether LIC have an intrinsic predilection for specific hematopoietic niches. For these purposes, lentiviral luciferase-transduced CD34-enriched and CD34-depleted cells from molecularly characterized buy Tedizolid (phosphate) samples were transplanted into neonatal RAG22/2cc2/2 mice that permit high levels of human hematopoietic engraftment. In this study, the CD34+ fraction of pediatric NOTCH1Mutated T-ALL samples had enhanced survival and self-renewal potential, characteristic of LIC, compared with their CD34+ NOTCH1 wild-type counterparts. These NOTCH1Mutated LIC were uniquely susceptible to targeted inhibition with a therapeutic human NOTCH1 monoclonal antibody selective for the NRR, while normal hematopoietic progenitors were spared thereby highlighting the cell type and context specific effects of NOTCH signaling and the importance of oncogenic addiction to NOTCH1 signaling in T-ALL LIC maintenance. Results T-ALL Molecular Characterization Molecular characterization of CD34+ cells from 12 T-ALL patient samples was performed by targeted exon sequencing analysis and focused on genes commonly mutated in T-ALL, including NOTCH1, PTEN, PIK3R1 and FBXW7. Selective NOTCH1 DNA sequencing revealed activating mutations in six of eleven newly diagnosed pediatric T-ALL samples and in one relapsed young adult T-ALL sample. In addition, CD34+ T-ALL cells derived from these 12 samples were further sequenced to identify PI3K, PTEN and FBXW7 pathway mutations common to pediatric T-ALL. Some cases harbored mutations in PTEN or PIK3R1 genes . PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/22200994 These data demonstrate that mutations in NOTCH1 and other 
genes capable of promoting LIC survival co-exist in the CD34+ fraction of T-ALL samples. NOTCH1Mutated T-ALL LIC are Serially Transplantable To determine if lentiviral luciferase-transduced CD34+ and CD342 cells from NOTCH1Mutated and NOTCH1WT pediatric TALL samples differed in their capacity to propagate disease, quantitative non-invasive bioluminescent imaging was performed within 10 weeks of intrahepatic transplantation of neonatal RAG22/2cc2/2 mice. Mice transplanted with CD34+ enriched NOTCH1Mutated T-ALL cells demonstrated significantly greater leukemic engraftment than mice transplanted with CD342 cells. Conversely, both CD34+ and CD342 fractions from NOTCH1WT T-ALL samples exhibited equivalent engraftment capacity in primary transplant recipients. Hence, CD34+ cells from NOTCH1Mutated samples gave rise to higher levels of bioluminescent engraftment in primary transplant recipients than their CD342 counterparts, indicative of LIC enrichment in the CD34+ fraction in NOTCH1Mutated but not NOTCH1WT samples. The predilection of NOTCH1Mutated T-ALL LIC for specific hematopoietic niches was determined in primary and serial transplants. Pr