The stem/progenitor landscape is reshaped in a mouse model of essential thrombocythemia and causes excess megakaryocyte production

Frameshift mutations in CALR (calreticulin) are associated with essential thrombocythemia (ET), but the stages at and mechanisms by which mutant CALR drives transformation remain incompletely defined. Here, we use single-cell approaches to examine the hematopoietic stem/progenitor cell landscape in a mouse model of mutant CALR-driven ET. We identify a trajectory linking hematopoietic stem cells (HSCs) with megakaryocytes and prospectively identify a previously unknown intermediate population that is overrepresented in the disease state. We also show that mutant CALR drives transformation primarily from the earliest stem cell compartment, with some contribution from megakaryocyte progenitors. Last, relative to wild-type HSCs, mutant CALR HSCs show increases in JAK-STAT signaling, the unfolded protein response, cell cycle, and a previously undescribed up-regulation of cholesterol biosynthesis. Overall, we have identified a novel megakaryocyte-biased cell population that is increased in a mouse model of ET and described transcriptomic changes linking CALR mutations to increased HSC proliferation and megakaryopoiesis

Two Aldehyde Clearance Systems Are Essential to Prevent Lethal Formaldehyde Accumulation in Mice and Humans.

Reactive aldehydes arise as by-products of metabolism and are normally cleared by multiple families of enzymes. We find that mice lacking two aldehyde detoxifying enzymes, mitochondrial ALDH2 and cytoplasmic ADH5, have greatly shortened lifespans and develop leukemia. Hematopoiesis is disrupted profoundly, with a reduction of hematopoietic stem cells and common lymphoid progenitors causing a severely depleted acquired immune system. We show that formaldehyde is a common substrate of ALDH2 and ADH5 and establish methods to quantify elevated blood formaldehyde and formaldehyde-DNA adducts in tissues. Bone-marrow-derived progenitors actively engage DNA repair but also imprint a formaldehyde-driven mutation signature similar to aging-associated human cancer mutation signatures. Furthermore, we identify analogous genetic defects in children causing a previously uncharacterized inherited bone marrow failure and pre-leukemic syndrome. Endogenous formaldehyde clearance alone is therefore critical for hematopoiesis and in limiting mutagenesis in somatic tissues

Preleukemic single-cell landscapes reveal mutation-specific mechanisms and gene programs predictive of AML patient outcomes

Acute myeloid leukemia (AML) and myeloid neoplasms develop through acquisition of somatic mutations that confer mutation-specific fitness advantages to hematopoietic stem and progenitor cells. However, our understanding of mutational effects remains limited to the resolution attainable within immunophenotypically and clinically accessible bulk cell populations. To decipher heterogeneous cellular fitness to preleukemic mutational perturbations, we performed single-cell RNA sequencing of eight different mouse models with driver mutations of myeloid malignancies, generating 269,048 single-cell profiles. Our analysis infers mutation-driven perturbations in cell abundance, cellular lineage fate, cellular metabolism, and gene expression at the continuous resolution, pinpointing cell populations with transcriptional alterations associated with differentiation bias. We further develop an 11-gene scoring system (Stem11) on the basis of preleukemic transcriptional signatures that predicts AML patient outcomes. Our results demonstrate that a single-cell-resolution deep characterization of preleukemic biology has the potential to enhance our understanding of AML heterogeneity and inform more effective risk stratification strategies

STAT1 is essential for HSC function and maintains MHCIIhi stem cells that resists myeloablation and neoplastic expansion

Adult hematopoietic stem cells (HSCs) are predominantly quiescent and can be activated in response to acute stress such as infection or cytotoxic insults. STAT1 is a pivotal downstream mediator of interferon (IFN) signaling and is required for IFN-induced HSC proliferation, but little is known about the role of STAT1 in regulating homeostatic hematopoietic stem/progenitor cells (HSPCs). Here we show that loss of STAT1 altered the steady state HSPC landscape, impaired HSC function in transplantation assays, delayed blood cell regeneration following myeloablation, and disrupted molecular programs which protect HSCs including control of quiescence. Our results also reveal STAT1-dependent functional HSC heterogeneity. A previously unrecognized subset of homeostatic HSCs with elevated MHCII expression (MHCIIhi) displayed molecular features of reduced cycling and apoptosis, and was refractory to 5-FU induced myeloablation. Conversely, MHCIIlo HSCs displayed increased megakaryocytic potential and were preferentially expanded in CALR mutant mice with thrombocytosis. Similar to mice, high MHCII expression is a feature of human HSCs residing in a deeper quiescent state. Our results therefore position STAT1 at the interface of stem cell heterogeneity and the interplay between stem cells and the adaptive immune system, areas of broad interest in the wider stem cell field

STAT1 is essential for HSC function and maintains MHCIIhi stem cells that resist myeloablation and neoplastic expansion.

Adult hematopoietic stem cells (HSCs) are predominantly quiescent and can be activated in response to acute stress such as infection or cytotoxic insults. STAT1 is a pivotal downstream mediator of interferon (IFN) signaling and is required for IFN-induced HSC proliferation, but little is known about the role of STAT1 in regulating homeostatic hematopoietic stem/progenitor cells (HSPCs). Here, we show that loss of STAT1 altered the steady state HSPC landscape, impaired HSC function in transplantation assays, delayed blood cell regeneration following myeloablation, and disrupted molecular programs that protect HSCs, including control of quiescence. Our results also reveal STAT1-dependent functional HSC heterogeneity. A previously unrecognized subset of homeostatic HSCs with elevated major histocompatibility complex class II (MHCII) expression (MHCIIhi) displayed molecular features of reduced cycling and apoptosis and was refractory to 5-fluorouracil-induced myeloablation. Conversely, MHCIIlo HSCs displayed increased megakaryocytic potential and were preferentially expanded in CALR mutant mice with thrombocytosis. Similar to mice, high MHCII expression is a feature of human HSCs residing in a deeper quiescent state. Our results therefore position STAT1 at the interface of stem cell heterogeneity and the interplay between stem cells and the adaptive immune system, areas of broad interest in the wider stem cell field

Late Quaternary environmental changes in Marguerite Bay, Antarctic Peninsula, inferred from lake sediments and raised beaches

The Antarctic Peninsula is one of the fastest-warming regions on Earth, but its palaeoenvironmental history south of 63° latitude is relatively poorly documented, relying principally on the marine geological record and short ice cores. In this paper, we present evidence of late-Quaternary environmental change from the Marguerite Bay region combining data from lake sediment records on Horseshoe Island and Pourquoi-Pas Island, and raised beaches at Horseshoe Island, Pourquoi-Pas Island and Calmette Bay. Lake sediments were radiocarbon dated and analysed using a combination of sedimentological, geochemical and microfossil methods. Raised beaches were surveyed and analysed for changes in clast composition, size and roundness. Results suggest a non-erosive glacial regime could have existed on Horseshoe Island from 35,780 (38,650–33,380) or 32,910 (34,630–31,370) cal yr BP onwards. There is radiocarbon and macrofossil evidence for possible local deglaciation events at 28,830 (29,370–28,320) cal yr BP, immediately post-dating Antarctic Isotopic Maximum 4, and 21,110 (21,510–20,730 interpolated) cal yr BP coinciding with, or immediately post-dating, Antarctic Isotopic Maximum 2. The Holocene deglaciation of Horseshoe Island commenced from 10,610 (11,000–10,300) cal yr BP at the same time as the early Holocene temperature maximum recorded in Antarctic ice cores. This was followed by the onset of marine sedimentation in The Narrows, Pourquoi-Pas Island, before 8850 (8480–9260) cal yr BP. Relative sea level high stands of 40.79 m above present at Pourquoi-Pas Island and 40.55 m above present at Calmette Bay occurred sometime after 9000 cal yr BP and suggest that a thicker ice sheet, including grounded ice streams, was present in this region of the Antarctic Peninsula than that recorded at sites further north. Isolation of the Narrows Lake basin on Pourquoi-Pas Island shows relative sea level in this region had fallen rapidly to 19.41 m by 7270 (7385–7155) cal yr BP. Chaetoceros resting spores suggest high productivity and stratified surface waters in The Narrows after 8850 (9260–8480) cal yr BP and beach clasts provide evidence of a period of increased wave energy at approximately 8000 yr BP. Lake sediment and beach data suggest an extended period of regional warming sometime between 6200 and 2030 cal yr BP followed by the onset of Neoglacial conditions from 2630 and 2030 cal yr BP in Narrows Lake and Col Lake 1, respectively. Diatom and δ<sup>13</sup>C vs C/N and macrofossil evidence suggest a potential increase in the number of birds and seals visiting the Narrows Lake catchment sometime after 2100 (2250–2000) cal yr BP, with enhanced nutrient enrichment evident after 1150 (1230–1080) cal yr BP, and particularly from c. 460 (540–380) cal yr BP. A very recent increase in Gomphonema species and organic carbon in the top centimetre of the Narrows Lake sediment core after c. 410 (490–320) cal yr BP, and increased sedimentation rates in the Col Lake 1 sediment core, after c. 400 (490–310) cal yr BP may be a response to the regional late-Holocene warming of the Antarctic Peninsula