Tissue stem cells often exhibit developmental stage-specific and sexually dimorphic properties, but the underlying mechanism remains largely elusive. By characterizing IGF1R signaling in hematopoietic cells, here we report that its disruption exerts sex-specific effects in adult hematopoietic stem and progenitor cells (HSPCs). Loss of IGF1R decreases the HSPC population in females but not in males, in part due to a reduction in HSPC proliferation induced by estrogen. In addition, the adult female microenvironment enhances engraftment of wild-type but not Igf1r-null HSPCs. In contrast, during gestation, when both female and male fetuses are exposed to placental estrogens, loss of IGF1R reduces the numbers of their fetal liver HSPCs regardless of sex. Collectively, these data support the interplay of IGF1R and estrogen pathways in HSPCs and suggest that the proliferation-promoting effect of estrogen on HSPCs is in part mediated via IGF1R signaling.

CRISPR is revolutionizing the ability to do somatic gene editing in mice for the purpose of creating new cancer models. Inactivation of theVHLtumor suppressor gene is the signature initiating event in the most common form of kidney cancer, clear cell renal cell carcinoma (ccRCC). Such tumors are usually driven by the excessive HIF2 activity that arises when theVHLgene product, pVHL, is defective. Given the pressing need for a robust immunocompetent mouse model of human ccRCC, we directly injected adenovirus-associated viruses (AAVs) encoding sgRNAs againstVHLand other known/suspected ccRCC tumor suppressor genes into the kidneys of C57BL/6 mice under conditions where Cas9 was under the control of one of two different kidney-specific promoters (Cdh16orPax8) to induce kidney tumors. An AAV targetingVhl, Pbrm1, Keap1, andTsc1reproducibly caused macroscopic ccRCCs that partially resembled human ccRCC tumors with respect to transcriptome and cell of origin and responded to a ccRCC standard-of-care agent, axitinib. Unfortunately, these tumors, like those produced by earlier genetically engineered mouse ccRCCs, are HIF2 independent.

The marker genes identified in endothelial cell clusters

The IMC antibody and nuclear staining list

Annotation of the cell clusters identified by IMC

Supplementary Figures with legend

Spearman Correlation Between NOTCH3 Expression and global genes, angiogenesis gene signature, and stroma_EMT_TGFbeta gene signature in TCGA and Collaboration dataset

The summary of clinical features of the cohort A patient samples for scRNA-seq study and cohort B patient samples for IMC

AbstractCancer immunotherapy has revolutionized the treatment of lung adenocarcinoma (LUAD); however, a significant proportion of patients do not respond. Recent transcriptomic studies to understand determinants of immunotherapy response have pinpointed stromal-mediated resistance mechanisms. To gain a better understanding of stromal biology at the cellular and molecular level in LUAD, we performed single-cell RNA sequencing of 256,379 cells, including 13,857 mesenchymal cells, from 9 treatment-naïve patients. Among the mesenchymal cell subsets, FAP+PDPN+ cancer-associated fibroblasts (CAF) and ACTA2+MCAM+ pericytes were enriched in tumors and differentiated from lung-resident fibroblasts. Imaging mass cytometry revealed that both subsets were topographically adjacent to the perivascular niche and had close spatial interactions with endothelial cells (EC). Modeling of ligand and receptor interactomes between mesenchymal and ECs identified that NOTCH signaling drives these cell-to-cell interactions in tumors, with pericytes and CAFs as the signal receivers and arterial and PLVAPhigh immature neovascular ECs as the signal senders. Either pharmacologically blocking NOTCH signaling or genetically depleting NOTCH3 levels in mesenchymal cells significantly reduced collagen production and suppressed cell invasion. Bulk RNA sequencing data demonstrated that NOTCH3 expression correlated with poor survival in stroma-rich patients and that a T cell–inflamed gene signature only predicted survival in patients with low NOTCH3. Collectively, this study provides valuable insights into the role of NOTCH3 in regulating tumor stroma biology, warranting further studies to elucidate the clinical implications of targeting NOTCH3 signaling.Significance:NOTCH3 signaling activates tumor-associated mesenchymal cells, increases collagen production, and augments cell invasion in lung adenocarcinoma, suggesting its critical role in remodeling tumor stroma.

The summary of gene signatures used in the study

ScRNA-seq gene expression of mesenchymal cells

Supplementary Figures 1-20, Table 1 from A Novel Androgen Receptor Splice Variant Is Up-regulated during Prostate Cancer Progression and Promotes Androgen Depletion–Resistant Growth

<p>Please note the Supplementary Methods were corrected by the authors and re-posted on March 16, 2009. The corrected file provides an updated GEO accession number. The original and revised versions are provided for reference.</p>

Supplementary Figures 1-5 from Compensatory Upregulation of Tyrosine Kinase Etk/BMX in Response to Androgen Deprivation Promotes Castration-Resistant Growth of Prostate Cancer Cells

Please note the Supplementary Methods were corrected by the authors and re-posted on March 16, 2009. The corrected file provides an updated GEO accession number. The original and revised versions are provided for reference.

Supplementary Figures: Figure S1. Syntenic regions between human chromosome 21q and mouse chromosome 8q. Figure S2. IHC staining of prostate lesions in Pb-Cre;T-ERG;PtenL/L and Pb-Cre;T-3Mb-Erg;PtenL/L mouse models. Figure S3. Overview of mouse prostate tissues microdissected for genomic DNA PCR analysis and microarray expression profiling. Figure S4. Cre-mediated deletion of the interstitial region in T-delta-Erg/Pten-null lesions. Figure S5. Additional data showing the EMT phenotype in prostate lesions developed in Pb-Cre;T-3Mb-Erg;PtenL/L mice. Figure S6. Expression analysis of AR target genes in T-∆-Erg/Pten-null and Ras/Pten prostate tumors. Figure S7. Expression analysis of select ETS factor genes in prostate tissues. Figure S8. Expression analysis of interstitial genes in T-delta-Erg/Pten-null versus Pten-null HG-PIN lesions.

Legends for Supplementary Figure S1-S8

Supplementary Data from Reverse Translating Molecular Determinants of Anti–Programmed Death 1 Immunotherapy Response in Mouse Syngeneic Tumor Models

Supplementary Table S2. Gene sets significantly (FDR q-val<0.25) enriched in T-delta-Erg/Pten-null HG-PINs compared to Pten-null HG-PINs.

Supplementary Table S1. Expression levels of select interstitial genes in HG-PIN lesions.