Modulation of protein methylation to improve hematopoietic stem cell engraftment
Modulation of protein methylation to improve hematopoietic stem cell engraftment
批准号:
10370387
负责人:
Kevin D Bunting
金额:
$31.2万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
2020
资助国家:
美国
项目状态:
未结题
起止时间:
2020-04-01 至 2025-03-31
关键词:
Active SitesAdultAllelesAreaBiochemicalBiochemistryBiological AssayBloodBlood CellsBone MarrowBone Marrow CellsBone Marrow CytometryBone Marrow TransplantationC-terminalCatalytic DomainCell CycleCell DeathCell MaintenanceCellsCellularityCessation of lifeComplexCre driverDataDefectDevelopmentEmbryoEngraftmentEquilibriumEtiologyExperimental HematologyFamilyFetal LiverFlow CytometryFunctional disorderFutureGenerationsHematological DiseaseHematologyHematopoiesisHematopoieticHematopoietic stem cellsHoloenzymesHomeostasisHomingInterventionKnock-outKnockout MiceLaboratoriesLeadLeucineLoxP-flanked alleleMeasuresMediatingMethylationMethyltransferaseMitogensModelingNatural regenerationOutputPathway interactionsPatternPhasePhosphoric Monoester HydrolasesPhosphorylationPhosphorylation SitePhosphotransferasesPlayPopulation DecreasesProceduresProcessProductionProtein KinaseProtein MethylationProtein Phosphatase 2A Regulatory Subunit PR53ProteinsPublic HealthRegulationRejuvenationResearchResearch PersonnelRoleRouteSafetySerineSignal PathwaySignal TransductionSpecificityStem cell transplantStressSupporting CellTNFSF5 geneTestingTherapeuticThreonineTimeUmbilical Cord Bloodbasebiological adaptation to stressclinical applicationconditional knockoutcytokineexperimental studyfitnesshematopoietic stem cell self-renewalhematopoietic transplantationimprovedknock-downmouse modelnoveloverexpressionpost-transplantpreventprotein phosphatase 6replication stressresponseself-renewalstem cell engraftmentstem cell homeostasissubcellular targeting
中文摘要
项目总结:
英文摘要
PROJECT SUMMARY:
Expansion of our understanding of hematopoiesis, including identification of key biochemical modulators, is
critical to the development of novel interventions and improved potentially curative procedures such as bone
marrow transplantation. While much study has occurred on the roles of kinases, the hematopoietic roles of the
serine/threonine, multi-subunit PP2A family phosphatases, PP2A and PP4, have only begun to be examined.
PP2Ac is reversibly methylated on its carboxy-terminal leucine (L309) by the methyltransferase, LCMT-1, and
the methylesterase, PME-1, and this methylation regulates formation of certain PP2A complexes, and thus
subcellular targeting and specificity. PP4, which is ~60% identical to PP2A, is also methylated on its carboxyl-
terminal leucine. We recently found LCMT-1 is also the PP4 methyltransferase and selectively regulates
certain PP4 complexes in addition to select PP2A complexes. Through the creation and initial study of a gene-
trap LCMT-1 knockout (KO) mouse and subsequently a pan-hematopoietic conditional knockout (cKO) of the
LCMT-1 allele, we show equally striking reductions in multilineage HSC competitive repopulation activity
despite relatively normal adult HSC homeostasis. Therefore, LCMT-1 may have cell intrinsic roles in promoting
HSC fitness uniquely during the stress at early times post-transplantation. This proposal is a multi-investigator
application that leverages the biochemistry and experimental hematology expertise of both PIs. Together we
have discovered that LCMT-1 plays a critical intrinsic role in hematopoiesis that is specific to HSCs in the post-
transplantation expansion, survival, and self-renewal phase. In contrast, LCMT1 doesn't play a role during
HSC maintenance and cycling during steady-state conditions. Because LCMT-1 substrates are limited to PP2A
family phosphatases due to the unique way LCMT-1 must interact with both specific active site and C-terminal
residues, we hypothesize that LCMT-1 promotes high levels of “methylation-dependent” PP2A and PP4
heterotrimers that in turn modulate key regulatory phosphorylation sites on critical proteins such as Raf to
promote mitogen-associated protein kinase (MAPK) pathway signaling required for the early proliferative drive
during hematopoietic repopulation. In this proposal, we will test this hypothesis and begin dissecting
mechanism by utilizing LCMT-1 cKO mice to define specific defects in adult HSC multilineage repopulating
activity using flow cytometry-based assays. Assays will compare steady-state and replicative stress responses.
The importance of LCMT-1 catalytic activity will be determined, the involvement of methylation-dependent
PP2A complexes in repopulating function will be evaluated, and therapeutic rejuvenation strategies will be
explored. Accomplishing these objectives will move the field forward by improving understanding of signaling
regulation of hematopoiesis and by providing avenues for manipulation of LCMT-1 and PP2A family
phosphatases to accelerate the rate of hematopoietic regeneration.
期刊论文(1)
专著(0)
科研奖励(0)
会议论文
DOI:
10.3390/antiox10060956
发表时间:
2021-06-15
期刊:
Antioxidants (Basel, Switzerland)
影响因子:
--
作者:
[Wang Z, Mi T, Bradley HL, Metts J, Sabnis H, Zhu W, Arbiser J, Bunting KD]
通讯作者:
Bunting KD
STAT5 Structure-Function in Hematopoiesis
-
批准号:7891083
-
项目类别:
-
资助金额:$10.0万
-
财政年份:2009
-
负责人:Kevin D Bunting
-
依托单位:
NHLBI Research Opportunities for Minority Students
-
批准号:7425797
-
项目类别:
-
资助金额:$9.24万
-
财政年份:2005
-
负责人:Kevin D Bunting
-
依托单位:
Role of TIMPs in Hematopoietic Stem Cell Biology
-
批准号:6766886
-
项目类别:
-
资助金额:$34.43万
-
财政年份:2003
-
负责人:Kevin D Bunting
-
依托单位:
Role of TIMPs in Hematopoietic Stem Cell Biology
-
批准号:7090866
-
项目类别:
-
资助金额:$33.62万
-
财政年份:2003
-
负责人:Kevin D Bunting
-
依托单位:
Role of TIMPs in Hematopoietic Stem Cell Biology
-
批准号:6663597
-
项目类别:
-
资助金额:$34.7万
-
财政年份:2003
-
负责人:Kevin D Bunting
-
依托单位:
Role of TIMPs in Hematopoietic Stem Cell Biology
-
批准号:6921335
-
项目类别:
-
资助金额:$34.43万
-
财政年份:2003
-
负责人:Kevin D Bunting
-
依托单位:
JAK/STAT Signaling in Hematopoietic Stem Cells
-
批准号:6829142
-
项目类别:
-
资助金额:$25.09万
-
财政年份:2002
-
负责人:Kevin D Bunting
-
依托单位:
STAT5 Structure-Function in Hematopoiesis
-
批准号:8133428
-
项目类别:
-
资助金额:$29.05万
-
财政年份:2002
-
负责人:Kevin D Bunting
-
依托单位:
JAK/STAT Signaling in Hematopoietic Stem Cells
-
批准号:7002741
-
项目类别:
-
资助金额:$24.5万
-
财政年份:2002
-
负责人:Kevin D Bunting
-
依托单位:
JAK/STAT Signaling in Hematopoietic Stem Cells
-
批准号:6908325
-
项目类别:
-
资助金额:$24.52万
-
财政年份:2002
-
负责人:Kevin D Bunting
-
依托单位:
Mechanisms of STAT5 function in hematopoietic stem and progenitor cells
-
批准号:8502966
-
项目类别:
-
资助金额:$33.93万
-
财政年份:2002
-
负责人:Kevin D Bunting
-
依托单位:
STAT5 Structure-Function in Hematopoiesis
-
批准号:7883524
-
项目类别:
-
资助金额:$29.35万
-
财政年份:2002
-
负责人:Kevin D Bunting
-
依托单位:
JAK/STAT Signaling in Hematopoietic Stem Cells
-
批准号:6707007
-
项目类别:
-
资助金额:$0.77万
-
财政年份:2002
-
负责人:Kevin D Bunting
-
依托单位:
Mechanisms of STAT5 function in hematopoietic stem and progenitor cells
-
批准号:9256459
-
项目类别:
-
资助金额:$33.93万
-
财政年份:2002
-
负责人:Kevin D Bunting
-
依托单位:
JAK/STAT Signaling in Hematopoietic Stem Cells
-
批准号:6621328
-
项目类别:
-
资助金额:$25.29万
-
财政年份:2002
-
负责人:Kevin D Bunting
-
依托单位:
Signaling Mechanisms in Stem Cell Plasticity
-
批准号:6896330
-
项目类别:
-
资助金额:$11.18万
-
财政年份:2002
-
负责人:Kevin D Bunting
-
依托单位:
Signaling Mechanisms in Stem Cell Plasticity
-
批准号:6637384
-
项目类别:
-
资助金额:$4.24万
-
财政年份:2002
-
负责人:Kevin D Bunting
-
依托单位:
JAK/STAT Signaling in Hematopoietic Stem Cells
-
批准号:6433897
-
项目类别:
-
资助金额:$25.29万
-
财政年份:2002
-
负责人:Kevin D Bunting
-
依托单位:
STAT5 Structure-Function in Hematopoiesis
-
批准号:7363253
-
项目类别:
-
资助金额:$30.03万
-
财政年份:2002
-
负责人:Kevin D Bunting
-
依托单位:
STAT5 Structure-Function in Hematopoiesis
-
批准号:7587511
-
项目类别:
-
资助金额:$30.03万
-
财政年份:2002
-
负责人:Kevin D Bunting
-
依托单位:
海外基金