• JAK inhibitors
• JAK–STAT pathway
• disease modification
• myelofibrosis
• targeted therapies

• Humans
• Primary Myelofibrosis/genetics
• Primary Myelofibrosis/drug therapy
• Primary Myelofibrosis/therapy
• Primary Myelofibrosis/metabolism
• Molecular Targeted Therapy/methods
• Janus Kinase 2/genetics
• Janus Kinase 2/antagonists & inhibitors
• Janus Kinase 2/metabolism
• Janus Kinase Inhibitors/therapeutic use
• Protein Kinase Inhibitors/therapeutic use
• Receptors, Thrombopoietin/genetics
• Calreticulin/genetics
• Calreticulin/antagonists & inhibitors
• Mutation
• Nitriles
• Signal Transduction/drug effects
• Pyrazoles/therapeutic use

• NET-2018-12365935 Ministero della Salute
• 21267 Associazione Italiana per la Ricerca sul Cancro
• ECS_00000017 National Recovery and Resilience Plan (NRRP)
• 2022F4WMR3 Ministero dell'Università e della Ricerca
• Ministero della Salute
• Associazione Italiana per la Ricerca sul Cancro
• Ministero dell’Università e della Ricerca

• AML
• JAK/STAT
• MPNs
• TPO
• c-MPL
• c-MPL isoforms

• Humans
• Protein Isoforms/genetics
• Protein Isoforms/metabolism
• Receptors, Thrombopoietin/genetics
• Receptors, Thrombopoietin/metabolism
• Animals
• Signal Transduction/genetics
• Hematopoiesis/genetics
• Thrombopoietin/metabolism
• Thrombopoietin/genetics

• 5/13/2018/NCD-III Indian Council of Medical Research
• YU/Seed grant/132-2022 Yenepoya University

• CDC5L
• FOXA1
• JAK2
• non-small cell lung cancer

• Humans
• Carcinoma, Non-Small-Cell Lung/genetics
• Carcinoma, Non-Small-Cell Lung/metabolism
• Up-Regulation/genetics
• S Phase
• Lung Neoplasms/genetics
• Lung Neoplasms/metabolism
• MAP Kinase Signaling System/genetics
• Cell Proliferation/genetics
• Gene Expression Regulation, Neoplastic
• Apoptosis/genetics
• MicroRNAs/genetics
• Cell Line, Tumor
• Cell Movement
• Hepatocyte Nuclear Factor 3-alpha/genetics
• Hepatocyte Nuclear Factor 3-alpha/metabolism
• Janus Kinase 2/genetics
• Janus Kinase 2/metabolism

• 201301091 The Health Department of Shanxi Province
• CZQM2022019 Young Talent Development Plan of Changzhou Health Commission
• CYF2022007 Health Young Scientific and Technological Talent Development Plan of Changzhou Medical Development Center and Changzhou Medical Association

• Myeloproliferative neoplasms
• myelofibrosis
• therapeutic approaches

• Apoptosis
• JAK2/STAT5
• MK-2206
• PI3K/AKT
• Ruxolitinib
• TNBC

Activation of compensatory signaling nodes in cancer often requires combination therapies that are frequently plagued by dose-limiting toxicities. Intestinal lymphatic drug absorption is seldom explored, although reduced toxicity and sustained drug levels would be anticipated to improve systemic bioavailability. A potent orally bioavailable multi-functional kinase inhibitor (LP-182) is described with intrinsic lymphatic partitioning for the combined targeting of phosphoinositide 3-kinase (PI3K) and mitogen-activated protein kinase (MAPK) signaling pathways without observable toxicity. We demonstrate selectivity and therapeutic efficacy through reduction of downstream kinase activation, amelioration of disease phenotypes, and improved survival in animal models of myelofibrosis. Our further characterization of synthetic and physiochemical properties for small molecule lymphatic uptake will support continued advancements in lymphatropic therapy for altering disease trajectories of a myriad of human disease indications.

Combination therapies simultaneously inhibiting different therapeutic targets in cancer is challenged by individual pharmacokinetic profiles. Here, the authors generate an orally provided multi-targeted kinase inhibitor that is lymphatic absorbed and increases survival in a murine model of myelofibrosis.

• Aurora A
• JAK2
• ROCK
• cell signaling
• drug resistance
• myeloproliferative neoplasms

• Calreticulin
• Essential thrombocythemia
• JAK inhibitors
• JAK2
• MPL/thrombopoietin receptor
• Myelofibrosis
• Myeloproliferative neoplasms
• Polycythemia Vera
• Targeted therapies
• Therapy

• Calreticulin/genetics
• Calreticulin/metabolism
• Humans
• Mutation
• Myeloproliferative Disorders/genetics
• Polycythemia Vera/genetics
• Polycythemia Vera/metabolism
• Thrombocythemia, Essential/genetics
• Thrombocythemia, Essential/metabolism
• Tumor Microenvironment

• CPI-0610
• Fedratinib
• Imetelstat
• JAK-STAT
• Myelofibrosis
• Pacritinib
• Ruxolitinib

• targeted therapies
• myeloproliferative disease

• JAK inhibitors
• Myelofibrosis
• chronic phase myelofibrosis
• clinical trial
• investigational drug
• non-jak inhibitors
• philadelphia negative myeloproliferative neoplasms

• Administration, Oral
• Adult
• Aged
• Aged, 80 and over
• Aminopyridines/administration & dosage
• Antineoplastic Combined Chemotherapy Protocols/pharmacokinetics
• Antineoplastic Combined Chemotherapy Protocols/therapeutic use
• Female
• Follow-Up Studies
• Humans
• Male
• Maximum Tolerated Dose
• Middle Aged
• Morpholines/administration & dosage
• Nitriles
• Primary Myelofibrosis/drug therapy
• Primary Myelofibrosis/pathology
• Prognosis
• Pyrazoles/administration & dosage
• Pyrimidines
• Tissue Distribution

• Combination therapy
• HDAC
• JAK2V617F
• Myeloproliferative neoplasms
• Ruxolitinib
• Vorinostat

• JAK2
• MPL/thrombopoietin receptor
• Myeloproliferative neoplasms
• calreticulin
• therapy

• ERK1/2 signaling
• JAK1/2 inhibition
• cell cycle
• inflammation
• myeloproliferative neoplasm

• allogeneic
• myelofibrosis
• relapse
• transplantation

• Clinical trials
• Gastric cancer
• Immunotherapy
• Molecular classification
• Targeted therapy

• Antineoplastic Agents/pharmacology
• Antineoplastic Agents/therapeutic use
• Biomarkers, Tumor/analysis
• Drug Resistance, Neoplasm/genetics
• Gastrectomy
• Genomics
• Humans
• Molecular Targeted Therapy/methods
• Neoplasm Recurrence, Local/genetics
• Neoplasm Recurrence, Local/mortality
• Neoplasm Recurrence, Local/pathology
• Neoplasm Recurrence, Local/therapy
• Patient Selection
• Precision Medicine/methods
• Quality of Life
• Stomach/pathology
• Stomach/surgery
• Stomach Neoplasms/genetics
• Stomach Neoplasms/mortality
• Stomach Neoplasms/pathology
• Stomach Neoplasms/therapy

• breast cancer
• chemotherapy-induced thrombocytopenia
• inflammatory cells
• recombinant human TPO

JAK inhibitors have been developed following the discovery of the JAK2V617F in 2005 as the driver mutation of the majority of non- BCR-ABL1 myeloproliferative neoplasms (MPNs). Subsequently, the search for JAK2 inhibitors continued with the discovery that the other driver mutations ( CALR and MPL) also exhibited persistent JAK2 activation. Several type I ATP-competitive JAK inhibitors with different specificities were assessed in clinical trials and exhibited minimal hematologic toxicity. Interestingly, these JAK inhibitors display potent anti-inflammatory activity. Thus, JAK inhibitors targeting preferentially JAK1 and JAK3 have been developed to treat inflammation, autoimmune diseases, and graft-versus-host disease. Ten years after the beginning of clinical trials, only two drugs have been approved by the US Food and Drug Administration: one JAK2/JAK1 inhibitor (ruxolitinib) in intermediate-2 and high-risk myelofibrosis and hydroxyurea-resistant or -intolerant polycythemia vera and one JAK1/JAK3 inhibitor (tofacitinib) in methotrexate-resistant rheumatoid arthritis. The non-approved compounds exhibited many off-target effects leading to neurological and gastrointestinal toxicities, as seen in clinical trials for MPNs. Ruxolitinib is a well-tolerated drug with mostly anti-inflammatory properties. Despite a weak effect on the cause of the disease itself in MPNs, it improves the clinical state of patients and increases survival in myelofibrosis. This limited effect is related to the fact that ruxolitinib, like the other type I JAK2 inhibitors, inhibits equally mutated and wild-type JAK2 (JAK2WT) and also the JAK2 oncogenic activation. Thus, other approaches need to be developed and could be based on either (1) the development of new inhibitors specifically targeting JAK2V617F or (2) the combination of the actual JAK2 inhibitors with other therapies, in particular with molecules targeting pathways downstream of JAK2 activation or the stability of JAK2 molecule. In contrast, the strong anti-inflammatory effects of the JAK inhibitors appear as a very promising therapeutic approach for many inflammatory and auto-immune diseases.

• JAK inhibitors
• MPN
• allosteric inhibitor
• auto-immune diseases
• inflammation

Along with molecular abnormalities (mutations in JAK2, Calreticulin (CALR) and MPL genes), chronic inflammation is the major hallmark of Myelofibrosis (MF). Here, we investigated the in vitro effects of crucial factors of the inflammatory microenvironment (Interleukin (IL)-1β, Tumor Necrosis Factor (TNF)-α, Tissue Inhibitor of Metalloproteinases (TIMP)-1 and ATP) on the functional behaviour of MF-derived circulating CD34⁺ cells.

We found that, regardless mutation status, IL-1β or TNF-α increases the survival of MF-derived CD34⁺ cells. In addition, along with stimulation of cell cycle progression to the S-phase, IL-1β or TNF-α ± TIMP-1 significantly stimulate(s) the in vitro clonogenic ability of CD34⁺ cells from JAK2^V617 mutated patients. Whereas in the JAK2^V617F mutated group, the addition of IL-1β or TNF-α + TIMP-1 decreased the erythroid compartment of the CALR mutated patients. Megakaryocyte progenitors were stimulated by IL-1β (JAK2^V617F mutated patients only) and inhibited by TNF-α. IL-1β + TNF-α + C-X-C motif chemokine 12 (CXCL12) ± TIMP-1 highly stimulates the in vitro migration of MF-derived CD34+ cells. Interestingly, after migration toward IL-1β + TNF-α + CXCL12 ± TIMP-1, CD34⁺ cells from JAK2^V617F mutated patients show increased clonogenic ability.

Here we demonstrate that the interplay of these inflammatory factors promotes and selects the circulating MF-derived CD34⁺ cells with higher proliferative activity, clonogenic potential and migration ability. Targeting these micro-environmental interactions may be a clinically relevant approach.

• circulating CD34+ cells
• inflammatory microenvironment
• migration
• myelofibrosis
• survival

A decade on from the description of JAK2 V617F, the MPNs are circumscribed by an increasingly intricate landscape. There is now evidence that they are likely the result of combined genetic dysregulation, with several mutated genes involved in the regulation of epigenetic mechanisms. Epigenetic changes are not due to a change in the DNA sequence but are reversible modifications that dictate the way in which genes may be expressed (or silenced). Among the epigenetic mechanisms, DNA methylation is probably the best described. Currently known MPN‐associated mutations now include JAK2, MPL, LNK, CBL, CALR, TET2, ASXL1, IDH1, IDH2, IKZF1 and EZH2. Enhancing our knowledge about the mutation profile of patients may allow them to be stratified into risk groups which would aid clinical decision making. Ongoing work will answer whether the use of epigenetic therapies as alterative pathway targets in combination with JAK inhibitors may be more effective than single agent treatment.

• DNA methylation
• cell signalling
• combination epigenetic therapy
• epigenetics
• histone modification
• janus kinase
• miRNA regulation
• myeloproliferative neoplasm

Inhibition of the constitutively activated JAK/STAT pathway in JAK2V617F mutated cells by the JAK1/JAK2 inhibitor ruxolitinib resulted in clinical benefits in patients with myeloproliferative neoplasms. However, evidence of disease-modifying effects remains scanty; furthermore, some patients do not respond adequately to ruxolitinib, or have transient responses, thus novel treatment strategies are needed. Here we demonstrate that ruxolitinib causes incomplete inhibition of STAT5 in JAK2V617F mutated cells due to persistence of phosphorylated serine residues of STAT5b, that conversely are targeted by PI3K and mTORC1 inhibitors. We found that PI3K/mTOR-dependent phosphorylation of STAT5b serine residues involves Protein Phosphatase 2A and its repressor CIP2A. The levels of CIP2A were found increased in cells harboring the JAK2V617F mutation, and we provide evidence of a correlation between clinical responses and the extent of CIP2A downregulation in myelofibrosis patients receiving the mTOR inhibitor RAD001 in a phase II clinical trial. To achieve maximal inhibition of STAT5 phosphorylation, we combined ruxolitinib with BKM120, a PI3K inhibitor, and RAD001, an mTOR inhibitor, obtaining improved efficacy in JAK2V617F mutated cell lines, primary patients’ cells, and JAK2V617F knock-in mice. These findings contribute to understanding the effectiveness of PI3K/mTOR inhibitors in MPN and argue for the rationale to develop combination clinical trials.

• MPN
• PI3K/mTOR inhibitors
• STAT5
• phosphorylation
• ruxolitinib

• Calreticulin
• JAK2V617F
• Myeloproliferative neoplasms
• TPO-R

• STAT3
• Stathmin 1
• myeloproliferative neoplasms
• paclitaxel
• ruxolitinib

• CXCR4
• JAK2 inhibitors
• hematopoiesis

• AKT
• In vivo
• MPL W515L
• MPL515/625/630
• Myeloproliferative neoplasms (MPN)
• STAT5

The classic BCR-ABL-negative myeloproliferative neoplasms (MPNs), a form of chronic malignant hemopathies, have been classified into polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF). ET and PMF are two similar disorders in their pathogenesis, which is marked by a key role of the megakaryocyte (MK) lineage. Whereas ET is characterized by MK proliferation, PMF is also associated with aberrant MK differentiation (myelodysplasia), leading to the release of cytokines in the marrow environment, which causes the development of myelofibrosis. Thus, PMF is associated with both myeloproliferation and different levels of myelodysplastic features. MPNs are mostly driven by mutated genes called MPN drivers, which abnormally activate the cytokine receptor/JAK2 pathway and their downstream effectors. The recent discovery of CALR mutations has closed a gap in our knowledge and has shown that this mutated endoplasmic reticulum chaperone activates the thrombopoietin receptor MPL and JAK2. These genetic studies have shown that there are two main types of MPNs: JAK2V617F-MPNs, including ET, PV, and PMF, and the MPL-/CALR-MPNs, which include only ET and PMF. These MPN driver mutations are associated with additional mutations in genes involved in epigenetics, splicing, and signaling, which can precede or follow the acquisition of MPN driver mutations. They are involved in clonal expansion or phenotypic changes or both, leading to myelofibrosis or leukemic transformation or both. Only a few patients with ET exhibit mutations in non-MPN drivers, whereas the great majority of patients with PMF harbor one or several mutations in these genes. However, the entire pathogenesis of ET and PMF may also depend on other factors, such as the patient’s constitutional genetics, the bone marrow microenvironment, the inflammatory response, and age. Recent advances allowed a better stratification of these diseases and new therapeutic approaches with the development of JAK2 inhibitors.

• Myeloprolifarative neoplasms
• myelofibrosis
• thrombocythemia

• Combination
• Myelofibrosis
• Myeloproliferative neoplasm
• Novel
• Targeted
• Therapy

• Animals
• Antineoplastic Agents/pharmacology
• Antineoplastic Agents/therapeutic use
• Antineoplastic Combined Chemotherapy Protocols/therapeutic use
• Epigenesis, Genetic/drug effects
• Humans
• Interferons/pharmacology
• Interferons/therapeutic use
• Janus Kinases/antagonists & inhibitors
• Molecular Targeted Therapy
• Myeloproliferative Disorders/diagnosis
• Myeloproliferative Disorders/genetics
• Myeloproliferative Disorders/metabolism
• Myeloproliferative Disorders/therapy
• Philadelphia Chromosome
• Protein Kinase Inhibitors/pharmacology
• Protein Kinase Inhibitors/therapeutic use
• Signal Transduction/drug effects

• R01 CA077816 NCI NIH HHS
• I01 CX000916 CSRD VA
• R01 CA155566 NCI NIH HHS
• CA77816 NCI NIH HHS
• P30 CA060553 NCI NIH HHS
• CA155566 NCI NIH HHS

• BCR-ABL1 negative myeloproliferative neoplasms
• CALR
• JAK2
• MPL
• apoptosis

• Apoptosis/genetics
• Fusion Proteins, bcr-abl/analysis
• Fusion Proteins, bcr-abl/genetics
• Humans
• Immunochemistry
• Mutation
• Myeloproliferative Disorders/drug therapy
• Myeloproliferative Disorders/genetics

• Animals
• Antineoplastic Agents/pharmacology
• Antineoplastic Agents/therapeutic use
• Genetic Variation
• Hematologic Neoplasms/drug therapy
• Hematologic Neoplasms/genetics
• Hematologic Neoplasms/metabolism
• Hematopoiesis/genetics
• Humans
• Janus Kinases/antagonists & inhibitors
• Janus Kinases/genetics
• Janus Kinases/metabolism
• Molecular Targeted Therapy
• Mutation
• Protein Kinase Inhibitors/pharmacology
• Protein Kinase Inhibitors/therapeutic use
• STAT Transcription Factors/genetics
• STAT Transcription Factors/metabolism
• Signal Transduction/drug effects

Inhibitors of JAK2 kinase are emerging as an important treatment modality for myeloproliferative neoplasms (MPN). However, similar to other kinase inhibitors, resistance to JAK2 inhibitors may eventually emerge through a variety of mechanisms. Effective drug combination is one way to enhance therapeutic efficacy and combat resistance against JAK2 inhibitors. To identify potential combination partners for JAK2 compounds in MPN cell lines, we performed pooled shRNA screen targeting 5,000 genes in the presence or absence of JAK2 blockade. One of the top hits identified was MYC, an oncogenic transcription factor that is difficult to inhibit directly, but could be targeted by modulation of upstream regulatory elements such as kinases. We demonstrate herein that PIM kinase inhibitors efficiently suppress MYC protein levels in MPN cell lines. Overexpression of MYC restores the viability of PIM inhibitor-treated cells, revealing causal relationship between MYC down-regulation and cell growth inhibition by PIM compounds. Combination of various PIM inhibitors with a JAK2 inhibitor results in significant synergistic growth inhibition of multiple MPN cancer cell lines and induction of apoptosis. Mechanistic studies revealed strong downregulation of phosphorylated forms of S6 and 4EBP1 by JAK2/PIM inhibitor combination treatment. Finally, such combination was effective in eradicating in vitro JAK2 inhibitor-resistant MPN clones, where MYC is consistently up-regulated. These findings demonstrate that simultaneous suppression of JAK2 and PIM kinase activity by small molecule inhibitors is more effective than either agent alone in suppressing MPN cell growth. Our data suggest that JAK2 and PIM combination might warrant further investigation for the treatment of JAK2-driven hematologic malignancies.

• Antineoplastic Agents/therapeutic use
• Humans
• Janus Kinase 2/antagonists & inhibitors
• Janus Kinase 2/metabolism
• Myeloproliferative Disorders/drug therapy
• Myeloproliferative Disorders/enzymology
• Practice Guidelines as Topic
• Protein Kinase Inhibitors/pharmacology
• Protein Kinase Inhibitors/therapeutic use
• Time Factors

The myeloproliferative neoplasms (MPN) are clonal myeloid disorders characterized by proliferation of mature myeloid cells, such that in polycythaemia vera (PV), the red cell proliferation dominates, platelets in essential thrombocythaemia (ET) and in myelofibrosis (MF), there may be cytopenia or proliferation, but the characteristic feature is the strikingly abnormal bone marrow stroma. These entities have a tendency to show phenotypic mimicry and may transform from one to another, for example, 20–30% of patients with PV are likely to develop MF. The significant event in this field was the recognition that Janus Kinase-2 (JAK2) activation was highly prevalent, followed by the description of the JAK2V617F mutation in 2005 (vide infra), which stimulated renewed interest in disease biology. Janus Kinase-2-targeted therapies have led to marked improvements for patients with this condition. However, it is obvious that the pathogenesis of these complex disorders reaches beyond this mutation; only 50–60% of patients with ET, for example, have the JAK2 mutation and several additional mutations have been described, which are of relevance in both the pathogenesis and clinical phenotype of these conditions.