Dichotomous regulation of lysosomes by MYC and TFEB controls hematopoietic stem cell fate

García‐Prat, Laura; Kaufmann, Kerstin B.; Schneiter, Florin; Voisin, Véronique; Murison, Alex; Chen, J.; Chan-Seng-Yue, Michelle; Gan, Olga I.; McLeod, Jessica; Smith, Sabrina; Shoong, Michelle; Parris, Darrien; Pan, Kristele; Zeng, Andy G.X.; Krivdova, Gabriela; Gupta, Kinam; Takayanagi, Shin‐ichiro; Wagenblast, Elvin; Wang, W.; Lupien, Mathieu; Schroeder, Timm; Xie, Stephanie Z.; Dick, John E.

doi:10.1101/2021.02.24.432720

Cited by 7 publications

(14 citation statements)

References 42 publications

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“…Next, BFP + GFP + PLAG1-S and c-MYC co-overexpressing Lin − CD34 + cells and their control counterparts were assessed for primitive cell maintenance ex vivo ( Figure 7B-C ). Consistent with other reports, 56,90-92 relative to control, ectopic c-MYC independently promotes hematopoietic differentiation, as measured by loss of CD34 + and gain of CD33 + cells ( Figure 7D-E , purple vs red). Over culture c-MYC OE cells also become enlarged relative to control ( Figure 7F , purple vs red) and display the highest rates of active translation ( Figure 7G ).…”

Section: Resultssupporting

confidence: 92%

“…(C) Change in variance-stabilizing transformed (vst) PLAG1 transcript expression in Lin − cord blood cells cultured for 2 or 4 days showing the P value from 1-tailed Student t -test 39 and in 72-hour cultured long-term (Lin − CD34 + CD38 − CD45RA − CD90 + CD49f + ) CB HSCs showing the P value from 1-tailed Student t -test and differential expression from DEseq analysis. 56 (D) Schematic of Lin − CD34 + CB HSPC in vitro and in vivo functional assay timelines and lentivectors used for overexpression of PLAG1 protein isoforms. (E) Primary CFU output by BFP + Lin − CD34 + cells overexpressing PLAG1-A, B, or S, or Luciferase control (n = 3 per experiment).…”

Section: Resultsmentioning

confidence: 99%

“… 93 Recent findings in cultured human HSPCs also establish that MYC drives their ex vivo activation via promotion of anabolic programs. 56 Therefore, we next investigated whether PLAG1-S acts through repression of MYC and whether MYC-mediated activation of translation could influence the ability of PLAG1-S to promote stemness.…”

Section: Resultsmentioning

confidence: 99%

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PLAG1 dampens protein synthesis to promote human hematopoietic stem cell self-renewal

Chahi

Belew

et al. 2022

Blood

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Hematopoietic stem cell (HSC) dormancy is understood as supportive of HSC function and their long-term integrity. While regulation of stress responses incurred as a result of HSC activation is recognized as important in maintaining stem cell function, little is understood of the preventative machinery present in human HSCs that may serve to resist their activation and promote HSC self-renewal. We demonstrate that the transcription factor PLAG1 is essential for long-term HSC function and when overexpressed endows a 15.6-fold enhancement in the frequency of functional HSC in stimulatory conditions. Genome-wide measures of chromatin occupancy and PLAG1-directed gene expression changes combined with functional measures reveal that PLAG1 dampens protein synthesis, restrains cell growth and division, and enhances survival, with the primitive cell advantages it imparts being attenuated by addition of the potent translation activator, c-MYC. We find PLAG1 capitalizes on multiple regulatory factors to ensure protective diminished protein synthesis including 4EBP1 and translation-targeting miR-127, and does so independently of stress response signaling. Overall, our study identifies PLAG1 as an enforcer of human HSC dormancy and self-renewal through its highly context-specific regulation of protein biosynthesis, and classifies PLAG1 among a rare set of bona fide regulators of mRNA translation in these cells. Our findings showcase the importance of regulated translation control underlying human HSC physiology, its dysregulation under activating demands, and the potential if its targeting for therapeutic benefit.

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Section: Resultssupporting

confidence: 92%

Section: Resultsmentioning

confidence: 99%

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PLAG1 dampens protein synthesis to promote human hematopoietic stem cell self-renewal

Chahi

Belew

et al. 2022

Blood

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“…As many of the upregulated gene sets following miR-130a OE were related to HSC programs, we performed GSEA using quiescent and activated HSC signatures (Forsberg et al, 2010) and observed enrichment of mitotic cell cycle activation and proliferation gene sets and depletion of quiescent genes with miR-130a OE (Figure 2F, S2E). We also observed significant enrichment of MYC targets following miR-130a OE (Figure 2F), consistent with HSC activation and quiescence exit (García-Prat et al, 2021; Laurenti et al, 2008). To determine whether miR-130a alters proliferation and cell cycle kinetics of HSC, we performed EdU incorporation assays utilizing fluorescently-labelled thymidine analog to measure nascent DNA synthesis in sorted LT-HSC and ST-HSC 3 days after transduction with control or miR-130a lentiviruses.…”

Section: Resultssupporting

confidence: 71%

Identification of the Global miR-130a Targetome Reveals a Novel Role for TBL1XR1 in Hematopoietic Stem Cell Self-Renewal and t(8;21) AML

Krivdova

Voisin

Schoof

et al. 2021

Preprint

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Gene expression profiling and proteome analysis of normal and malignant hematopoietic stem cells (HSC) point to shared core stemness properties. However, discordance between mRNA and protein signatures underscores an important role for post-transcriptional regulation by miRNAs in governing this critical nexus. Here, we identified miR-130a as a regulator of HSC self-renewal and differentiation. Enforced expression of miR-130a impaired B lymphoid differentiation and expanded long-term HSC. Integration of protein mass spectrometry and chimeric AGO2 eCLIP-seq identified TBL1XR1 as a primary miR-130a target, whose loss of function phenocopied miR-130a overexpression. Moreover, we found that miR-130a is highly expressed in t(8;21) AML where it is critical for maintaining the oncogenic molecular program mediated by AML1-ETO. Our study establishes that identification of the comprehensive miRNA targetome within primary cells enables discovery of novel genes and molecular networks underpinning stemness properties of normal and leukemic cells.

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“…Further supporting the molecular features of dormant cells, the IBα KO LT-HSCs also express significantly higher levels of the transcription factor EB (tfeb) (Figure 4F), whose elevated levels is a further indicator of dormant cell (García-Prat et al, 2021;Garcia Prat et al, 2020).…”

Section: Increased Activation Of Retinoic Acid Receptor Rarα In the Agmmentioning

confidence: 64%

IκBα controls dormancy induction in Hematopoietic stem cell development via retinoic acid

Thambyrajah

Fadlullah

Proffitt

et al. 2022

Preprint

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Recent findings are challenging the classical hematopoietic model in which long-term hematopoietic stem cells (LT-HSC) are the base of the hematopoietic system. Clonal dynamics analysis of the hematopoietic system indicate that LT-HSC are not the main contributors of normal hemapoiesis in physiological conditions and the hematopoietic system is mainly maintained by multipotent progenitors (MPPs, hereafter HPC) and LT-HSCs are mostly in a non-active state. The first HSCs emerge from the aorta-gonad and mesonephros (AGM) region along with hematopoietic progenitors (HPC) within hematopoietic clusters. Molecular pathways that determine the HSC fate instead of HPC are still unknown, although inflammatory signaling, including NF-ΚB has been implicated in the development of HSCs. Here, we identify a chromatin binding function for IΚBα (also known as the inhibitor of NF-ΚB) that is Polycomb repression complex 2 (PRC2)- dependent and specifically determines dormant vs proliferating HSCs from the onset of their emergence in the AGM. We find a specific reduction of LT-HSCs in the IΚBα knockout new-born pups. This defect is manifested at the FL stage already, and traceable to the first emerging HSCs in the E11.5 AGM, without affecting the general HPC population. IΚBα-deficient LT-HSCs express dormancy signature genes, are less proliferative and can robustly respond to activation stimuli such asin vitroculture and serial transplantation. At the molecular level, we find decreased PRC2-dependent H3K27me3 at the promoters of several retinoic acid signaling elements in the IΚBα- deficient aortic endothelium and E14.5 FL LT-HSCs. Additionally, IΚBα binding itself is found in the promoters of retinoic acid receptors rarα in the AGM, and rarγ in the LT-HSC of FL. Overall, we demonstrate that the retinoic acid pathway is over-activated in the hematopoietic clusters of IΚBα-deficient AGMs leading to premature dormancy of LT- HSCs that persists in the FL LT-HSCs.

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Dichotomous regulation of lysosomes by MYC and TFEB controls hematopoietic stem cell fate

Cited by 7 publications

References 42 publications

PLAG1 dampens protein synthesis to promote human hematopoietic stem cell self-renewal

PLAG1 dampens protein synthesis to promote human hematopoietic stem cell self-renewal

Identification of the Global miR-130a Targetome Reveals a Novel Role for TBL1XR1 in Hematopoietic Stem Cell Self-Renewal and t(8;21) AML

IκBα controls dormancy induction in Hematopoietic stem cell development via retinoic acid

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