PMID- 35953207
OWN - NLM
STAT- MEDLINE
DCOM- 20220815
LR  - 20220823
IS  - 0091-679X (Print)
IS  - 0091-679X (Linking)
VI  - 171
DP  - 2022
TI  - Immunoblotting-assisted assessment of JAK/STAT and PI3K/Akt/mTOR signaling in 
      myeloproliferative neoplasms CD34+ stem cells.
PG  - 81-109
LID - S0091-679X(22)00052-8 [pii]
LID - 10.1016/bs.mcb.2022.04.005 [doi]
AB  - Philadelphia-negative myeloproliferative neoplasms (pH-MPNs) origin from the 
      clonal expansion of hematopoietic stem cells with acquired mutations leading to 
      uncontrolled proliferation of differentiated myeloid cells. The main entities of 
      Ph-MPNs are represented by Polycythemia Vera (PV), Essential Thrombocythemia (ET) 
      and Myelofibrosis (MF) that are characterized by microvascular disorders, 
      thrombosis and bleeding, splenomegaly secondary to extramedullary hematopoiesis, 
      various degree of bone marrow fibrosis and a progressive risk of leukemic 
      transformation. Somatic mutations in myeloid genes including JAK2, CALR, and MPL 
      cause the constitutive activation of the Janus Kinase 2 (JAK)/signal transducer 
      and activator of transcription (STAT) pathway that confers proliferative and 
      differentiative advantage to mutated hematopoietic progenitors and ultimately 
      drives the development of a Ph-MPNs phenotype. Beyond the JAK/STAT axis, a wide 
      number of intracellular signaling pathways were found deregulated in Ph-MPNs 
      including the phosphatidylinositol-3-kinase (PI3K)/protein kinase B 
      (Akt)/mammalian target of rapamycin (mTOR) constitutive activation. In this 
      chapter, we provide a detailed protocol for the immunoblotting assisted 
      assessment of Ph-MPNs pathways activation. This protocol can be easily adapted to 
      study protein expression and phosphorylation of hematopoietic stem progenitors 
      and differentiated cell lineages.
CI  - Copyright (c) 2022 Elsevier Inc. All rights reserved.
FAU - Calabresi, Laura
AU  - Calabresi L
AD  - Center for Research and Innovation for Myeloproliferative Neoplasms, Department 
      of Experimental and Clinical Medicine, University of Florence, Florence, Italy.
FAU - Balliu, Manjola
AU  - Balliu M
AD  - Center for Research and Innovation for Myeloproliferative Neoplasms, Department 
      of Experimental and Clinical Medicine, University of Florence, Florence, Italy.
FAU - Bartalucci, Niccolo
AU  - Bartalucci N
AD  - Center for Research and Innovation for Myeloproliferative Neoplasms, Department 
      of Experimental and Clinical Medicine, University of Florence, Florence, Italy. 
      Electronic address: niccolo.bartalucci@unifi.it.
LA  - eng
PT  - Journal Article
DEP - 20220726
PL  - United States
TA  - Methods Cell Biol
JT  - Methods in cell biology
JID - 0373334
RN  - 0 (Calreticulin)
RN  - EC 2.7.1.1 (MTOR protein, human)
RN  - EC 2.7.10.2 (Janus Kinase 2)
RN  - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt)
RN  - EC 2.7.11.1 (TOR Serine-Threonine Kinases)
SB  - IM
MH  - Calreticulin/genetics
MH  - Humans
MH  - Immunoblotting
MH  - Janus Kinase 2/genetics
MH  - Mutation
MH  - *Myeloproliferative Disorders/genetics
MH  - Phosphatidylinositol 3-Kinases/genetics
MH  - *Polycythemia Vera/genetics
MH  - *Primary Myelofibrosis/genetics
MH  - Proto-Oncogene Proteins c-akt/genetics
MH  - Stem Cells
MH  - TOR Serine-Threonine Kinases/genetics
OTO - NOTNLM
OT  - CD34+ cells
OT  - Essential thrombocythemia
OT  - Hematopoietic colonies
OT  - Immunoblotting
OT  - JAK2
OT  - Myelofibrosis
OT  - Myeloproliferative neoplasms
OT  - Polycythemia vera
OT  - STAT5
COIS- Disclosures LC, MB, and NB have no conflicts of interest to disclose.
EDAT- 2022/08/12 06:00
MHDA- 2022/08/16 06:00
CRDT- 2022/08/11 21:06
PHST- 2022/08/11 21:06 [entrez]
PHST- 2022/08/12 06:00 [pubmed]
PHST- 2022/08/16 06:00 [medline]
AID - S0091-679X(22)00052-8 [pii]
AID - 10.1016/bs.mcb.2022.04.005 [doi]
PST - ppublish
SO  - Methods Cell Biol. 2022;171:81-109. doi: 10.1016/bs.mcb.2022.04.005. Epub 2022 
      Jul 26.