Work Package (WP)
WP1: Hematopoiesis stem cells (HSCs) changes during ageing [Months: 8-44] UMCGBeneficiary: UMCG, CERBM-GIE, FM EU S.P.A. and MPG
The main focuses of this WP are the physiological age-related changes in HSCs properties and the identification of key genes involved in this process. The WP consists of the following tasks: T1: improve protocols to work with low number of cells (UMCG, CERBM, MPG); T2: compare young and old HSCs in different mouse strains by RNA-seq profiling, MeDIP analysis, ChIP-seq analysis (UMCG). T3: define physiological HSCs properties of young vs old HSCs by serial transplantation of virally barcoded HSCs (UMCG). These approaches will compare the molecular profile (at the transcriptome and epigenome levels) of HSCs from different mouse strains and of different ages (UMCG). T4: define the role of Helios (IKZF2) in controlling the age-dependent evolution of transcriptional and epigenetic landscapes in HSCs and its impact on hematopoietic cell fitness. This will be studied with a particular focus on genome stability (CERBM). T5: define epigenomic and transcription factor occupancy maps in DNA repair defective (Ercc1-/-) aged HSCs and their differentiation potential (FORTH). T6-T8: get insight into the potential transcriptional functions of DNA repair proteins in HSC aging (FORTH). T9: improve polychromatic flow cytometry for the detection of intracellular and cell surface biomarkers to create reference profile panels for profiling young vs adult blood cells populations (FME, BD). T10: to address the evolution of the dormant compartment from young towards aged hematopoiesis (MPG). T11: address abilityof single aged dormant HSCs to return to quiescence upon stress (MPG).
WP2: Genetic and epigenetic control of transcription [Months: 8-44] SAPIENZABeneficiary: UNIMIB, CERBM-GIE, SAPIENZA, Vetmeduni Vienna and CSIC
The main question addressed in this WP is how hematopoietic cell identity is controlled at the transcriptional and epigenetic levels in normal hematopoiesis and in leukemias. The WP consists of the following tasks: T1: Identification effector candidates of AML oncoproteins and their functional characterization by loss-of-function experiments (VETMEDUNI); T2: identification of Sox6 and NR2F2 regulated networks in erythropoiesis and functional characterization by loss-of-function/overexpression of selected downstream genes (UNIMIB); T3: deciphering aberrant epigenetic marks in AML cells by proteomic analysis and lncRNA profiling (CRCM); T4: study of the molecular mechanisms underlying the formation polycomb-repressive islets during leukemogenesis (CRCM); T5: characterization of non-malignant and malignant transformation of hematopoietic progenitors carrying alleles for inducible inactivation of PRC1 subunits (CSIC); T6: comparison of the transcriptome profiles of stable self-renewing multipotent/oligopotent progenitors and of immature lineage-committed precursors (CSIC); T7: investigation of the role of m6A modification in AML cells, by knocking down writers, erasers and readers of m6A modification (UNIROMA1); T8: purification of the components of the m6A writer complex to define their potential role in AML (UNIROMA1). The work in WP2 will take advantage of technologies developed by FME and DIAGENODE.
WP3:Intrinsic and niche-dependent signalling in acute leukemias [Months: 8-44] INSTITUT CURIEBeneficiary: INSTITUT CURIE, TAU, FT, DIAGENODE and BRFAA
This WP aims to understand the crosstalk between intrinsic and extrinsic cues that sustain the proliferation of preleukemic and leukemic cells within the hematopoietic niche. It consists of the following tasks. Based on IC data that T-ALL maintenance requires CXCR4 activation by stroma-derived CXCL12 (Cancer Cell 2015, 27:769-79) while persistent re-activation of the TCR is anti-leukemic in vivo (Cancer Discov. 2016,6:972-85), IC will: T1. investigate the biological/therapeutic response of T-ALL xenografts to monotherapy using CXCR4 and CXCL12 inhibitors; T2. dissect anti-CD3e-induced cell death pathways in T-ALL; T3. investigate therapeutic synergy between CXCR4/CXCL12 inhibitors or anti-CD3e mAb with a conventional chemotherapy regimen (vincristine+dexamethasone). SCHNEIDER CHILDREN’S will: T4. model the involvement of thymic stromal lymphopoietin (TSLP) receptor and TSLP in ALL in vitro (grown on stromal cells) and ALL xenografts; T5. investigate the role of of hTSLP in growth and drug resistance of CRLF2 (TSLPR) ALL, by growing primary ALL cells on MS5-hTSLP stroma and test the efficacy of anti-TSLP and anti-TSLPR drugs on in vivo and in vitro growth of engineered preleukemia cells and primary ALL cells. FT will: T6. generate an in vivo model of the preleukemic niche by heterotopic transplantation of preleukemic cells in immunocompromised mice; T7. investigate whether the niche provided by human heterotopic stem cells niche can promote the evolution from pre-leukemia to leukemias; T8. DIAGENODE will develop epigenomics to identify variable regions that account for MLL- rearranged-AML specificity, ii) evaluate the impact of epigenomic variations on disease characteristics by confronting epigenomics and clinical data; T9. BRFAA will use MDS and AML, as models of preleukemia and leukemia, to investigate how STAT5 target gene networks change in leukemia progression.
WP4: Structured training program (Training) [Months: 8-44] UNIMIBBeneficiary: UNIMIB, UMCG, CERBM-GIE, KCL, FM EU S.P.A., SAPIENZA, Vetmeduni Vienna, MPG, INSERM, CSIC, INSTITUT CURIE, TAU, FT, DIAGENODE and BRFAA
The ARCH training programme is organized to provide ESRs with a solid scientific and technological knowledge, together with a portfolio of transferable skills that will integrate their training and will improve their future careers prospects. WP5 will involve the active participation of partners, who will make their specific expertise available to ARCH ESRs. The WP consists of the following tasks. T1: training through research: single research projects, including secondments to collaborating academic laboratories and private partners; T2: definition of a Personal Development Career plans (see also WP6) and enrollment of ESRs in academic PhD programmes; T3: organization of local training activities (LTM, 1.2.1 a) and monitoring the ESRs participation; T4: organization of ARCH Network-wide training events (NTM, 1.2.1 b and Table 1.2.b); T5: ensuring a proper ESRs’ participation in international conferences/events (International external training module IETM, 1.2.1 c); T6: guiding and advising students to plan their career after ARCH completion.