Single-cell transcriptomics uncovers distinct molecular signatures of stem cells in chronic myeloid leukemia

Recent advances in single-cell transcriptomics are ideally placed to unravel intratumoral heterogeneity and selective resistance of cancer stem cell (SC) subpopulations to molecularly targeted cancer therapies. However, current single-cell RNA-sequencing approaches lack the sensitivity required to reliably detect somatic mutations. We developed a method that combines high-sensitivity mutation detection with whole-transcriptome analysis of the same single cell. We applied this technique to analyze more than 2,000 SCs from patients with chronic myeloid leukemia (CML) throughout the disease course, revealing heterogeneity of CML-SCs, including the identification of a subgroup of CML-SCs with a distinct molecular signature that selectively persisted during prolonged therapy. Analysis of nonleukemic SCs from patients with CML also provided new insights into cell-extrinsic disruption of hematopoiesis in CML associated with clinical outcome. Furthermore, we used this single-cell approach to identify a blast-crisis-specific SC population, which was also present in a subclone of CML-SCs during the chronic phase in a patient who subsequently developed blast crisis. This approach, which might be broadly applied to any malignancy, illustrates how single-cell analysis can identify subpopulations of therapy-resistant SCs that are not apparent through cell-population analysis

Single-cell transcriptomics uncovers distinct molecular signatures of stem cells in chronic myeloid leukemia

Recent advances in single-cell transcriptomics are ideally placed to unravel intratumoral heterogeneity and selective resistance of cancer stem cell (SC) subpopulations to molecularly targeted cancer therapies. However, current single-cell RNA-sequencing approaches lack the sensitivity required to reliably detect somatic mutations. We developed a method that combines high-sensitivity mutation detection with whole-transcriptome analysis of the same single cell. We applied this technique to analyze more than 2,000 SCs from patients with chronic myeloid leukemia (CML) throughout the disease course, revealing heterogeneity of CML-SCs, including the identification of a subgroup of CML-SCs with a distinct molecular signature that selectively persisted during prolonged therapy. Analysis of nonleukemic SCs from patients with CML also provided new insights into cell-extrinsic disruption of hematopoiesis in CML associated with clinical outcome. Furthermore, we used this single-cell approach to identify a blast-crisis-specific SC population, which was also present in a subclone of CML-SCs during the chronic phase in a patient who subsequently developed blast crisis. This approach, which might be broadly applied to any malignancy, illustrates how single-cell analysis can identify subpopulations of therapy-resistant SCs that are not apparent through cell-population analysis

First Scattered-light Images of the Gas-rich Debris Disk around 49 Ceti

We present the first scattered-light images of the debris disk around 49 Ceti, a ∼40 Myr A1 main-sequence star at 59 pc, famous for hosting two massive dust belts as well as large quantities of atomic and molecular gas. The outer disk is revealed in reprocessed archival Hubble Space Telescope NICMOS-F110W images, as well as new coronagraphic H-band images from the Very Large Telescope SPHERE instrument. The disk extends from 1.″1 (65 au) to 4.″6 (250 au) and is seen at an inclination of 73°, which refines previous measurements at lower angular resolution. We also report no companion detection larger than 3 M Jup at projected separations beyond 20 au from the star (0.″34). Comparison between the F110W and H-band images is consistent with a gray color of 49 Ceti's dust, indicating grains larger than 2 μm. Our photometric measurements indicate a scattering efficiency/infrared excess ratio of 0.2-0.4, relatively low compared to other characterized debris disks. We find that 49 Ceti presents morphological and scattering properties very similar to the gas-rich HD 131835 system. From our constraint on the disk inclination we find that the atomic gas previously detected in absorption must extend to the inner disk, and that the latter must be depleted of CO gas. Building on previous studies, we propose a schematic view of the system describing the dust and gas structure around 49 Ceti and hypothetical scenarios for the gas nature and origin.E.C. acknowledges support from NASA through Hubble Fellowship grant HST-HF2-51355 awarded by STScI, operated by AURA, Inc. under contract NAS5-26555, and support from HST-AR-12652, for research carried out at the Jet Propulsion Laboratory, California Institute of Technology. J.M. acknowledges ESO through the ESO fellowship program. M.B. acknowledges support from DFG project Kr 2164/15-1. G.M.K. is supported by the Royal Society as a Royal Society University Research Fellow. C.d.B. is supported by Mexican CONACyT research grant CB-2012-183007. L.M. acknowledges support by STFC through a graduate studentship. J.C.A. acknowledges support by the Programme National de Planétologie. We acknowledge support by the European Union through ERC grant 337569 for O.A. and C.A.G.G. and grant 279973 for M.W. and L.M

Origins of ETP Leukemia

The relationship between the identity of the normal cells targeted by oncogenic mutations and the phenotype of the resulting cancer is not well understood. In a recent study [1], we sought to address this question by modelling a distinct and chemotherapy-resistant subtype of T cell acute lymphoblastic leukemia (T-ALL), known as early thymic progenitor (ETP) leukemia

Cell-extrinsic hematopoietic impact of Ezh2 inactivation in fetal liver endothelial cells

Despite the well-established cell-intrinsic role of epigenetic factors in normal and malignant hematopoiesis, their cell-extrinsic role remains largely unexplored. Herein we investigated the hematopoietic impact of inactivating Ezh2, a key component of polycomb repressive complex 2 (PRC2), in the fetal liver (FL) vascular niche. Hematopoietic specific (Vav-iCre) Ezh2-inactivation enhanced FL hematopoietic stem cell (HSC) expansion with normal FL erythropoiesis. In contrast, endothelium (Tie2-Cre) targeted Ezh2-inactivation resulted in embryonic lethality with severe anemia at E13.5 despite normal emergence of functional HSCs. Ezh2-deficient FL endothelium overexpressed Mmp9 which cell-extrinsically depleted the membrane-bound form of Kit ligand (mKitL), an essential hematopoietic cytokine, in FL. Furthermore, Mmp9 inhibition in vitro restored mKitL expression along with the erythropoiesis supporting capacity of FL endothelial cells. These data establish that Ezh2 is intrinsically dispensable for FL HSCs and provides proof of principle that modulation of epigenetic regulators in niche components can exert a marked cell-extrinsic impact

Cell-extrinsic hematopoietic impact of Ezh2 inactivation in fetal liver endothelial cells

Despite the well-established cell-intrinsic role of epigenetic factors in normal and malignant hematopoiesis, their cell-extrinsic role remains largely unexplored. Herein we investigated the hematopoietic impact of inactivating Ezh2, a key component of polycomb repressive complex 2 (PRC2), in the fetal liver (FL) vascular niche. Hematopoietic specific (Vav-iCre) Ezh2-inactivation enhanced FL hematopoietic stem cell (HSC) expansion with normal FL erythropoiesis. In contrast, endothelium (Tie2-Cre) targeted Ezh2-inactivation resulted in embryonic lethality with severe anemia at E13.5 despite normal emergence of functional HSCs. Ezh2-deficient FL endothelium overexpressed Mmp9 which cell-extrinsically depleted the membrane-bound form of Kit ligand (mKitL), an essential hematopoietic cytokine, in FL. Furthermore, Mmp9 inhibition in vitro restored mKitL expression along with the erythropoiesis supporting capacity of FL endothelial cells. These data establish that Ezh2 is intrinsically dispensable for FL HSCs and provides proof of principle that modulation of epigenetic regulators in niche components can exert a marked cell-extrinsic impact