miR-199b, a novel tumor suppressor miRNA in acute myeloid leukemia with prognostic implications

Additional file 1: Figure S1. miR-199b-5p targets HIF-1 alpha. Transcript levels of three predicted targets of miR-199b were tested via RT-qPCR in miR-199b silenced CD34 cells and HIF-1a levels were significantly increased by anti-miR-199b

Cross-Talk Between JNK/SAPK and ERK/MAPK Pathways: Sustained Activation of JNK Blocks ERK Activation by Mitogenic Factors

Mixed lineage kinases (MLKs) are a family of serine/threonine kinases that function in the SAPK signaling cascade. MLKs activate JNK/SAPK in vivo by directly phosphorylating and activating the JNK kinase SEK-1 (MKK4 and -7). Importantly, the MLK member MLK3/SPRK has been shown recently to be a direct target of ceramide and tumor necrosis factor-α (TNF-α) and to mediate the TNF-α and ceramide-induced JNK activation in Jurkat cells. Here we report that MLK3 can phosphorylate and activate MEK-1 directly in vitro and also can induce MEK phosphorylation on its activation sites in vivo in COS-7 cells. Surprisingly, this induction of MEK phosphorylation does not result in ERK activation in vivo. Rather, in cells expressing active MLK3, ERK becomes resistant to activation by growth factors and mitogens. This restriction in ERK activation requires MLK3 kinase activity, is independent of Raf activation, and is reversed by JNK pathway inhibition either at the level of SEK-1, JNK, or Jun. These results demonstrate that sustained JNK activation uncouples ERK activation from MEK in a manner requiring Jun-mediated gene transcription. This in turn points to the existence of a negative cross-talk relationship between the stress-activated JNK pathway and the mitogen-activated ERK pathway. Thus, our findings imply that some of the biological functions of JNK activators, such as TNF-α and ceramide, may be attributed to their ability to block cell responses to growth and survival factors acting through the ERK/MAPK pathway

Low-cost control of discontinuous systems including impacts and friction

For a successful design of an engineering system it is essential to pay careful attention to its dynamic response. This is particularly true, in the case of nonlinear systems, since they can exhibit very complex dynamic behaviour, including multiple co-existing stable solutions and chaotic motions, characterized by large sensitivity to initial conditions. In some systems nonlinear characteristics are desired and designed for, but in other cases they are unwanted and can cause fatigue and failure. A type of dynamical system which is highly nonlinear is discontinuous or non-smooth systems. In this work, systems with impacts are primarily investigated, and this is a typical example of a discontinuous system. To enhance or optimize the performance of dynamical systems, some kind of control can be implemented. This thesis concerns implementation of low-cost control strategies for discontinuous systems. Low-cost control means that a minimum amount of energy is used when performing the control actions, which is a desirable situation regardless of the application. The disadvantage of such a method is that the performance might be limited as compared with a control strategy with no restrictions on energy consumption. In this work, the control objective is to enforce a continuous or discontinuous grazing bifurcation of the system, whichever is desirable. In Paper A, the dynamic response and bifurcation behaviour of an impactoscillator with dry friction is investigated. For a one-degree-of-freedom model of the system, analytical solutions are found in separate regions of state space. These are then used to perform a perturbation analysis around a grazing trajectory. Through the analysis, a condition on the parameters of the system is derived, which assures a continuous grazing bifurcation. It is also shown that the result has bearing on the dynamic response of a two-degree-of-freedom model of the system. A low-cost active control strategy for a class of impact oscillators is proposed in Paper B. The idea of the control method is to introduce small adjustments in the position of the impact surface, at discrete moments in time, to assure a continuous bifurcation. A proof is given for what control parameters assures the stabilization. In Paper C, the proposed low-cost control method is implemented in a quarter-car model of a vehicle suspension, in order to minimize impact velocities with the bumpstop in case of high amplitude excitation. It is shown that the control method is effective for harmonic road excitation.QC 2010111

MicroRNA199b regulates mouse hematopoietic stem cells maintenance

The preservation of hematopoieticstem cell pool in bone marrow (BM) is crucial for sustained hematopoiesisin adults. Studies assessing adult hematopoieticstem cells functionality had been shown that for example loss of quiescence impairs hematopoieticstem cells maintenance. Although, miR‐199b is frequently down‐regulated in acute myeloid leukemia, its role in hematopoieticstem cells quiescence, self‐renewal and differentiation is poorly understood. Our laboratory investigated the role of miR‐199b in hematopoieticstem and progenitor cells (HSPCs) fate using miR‐199b‐5p global deletion mouse model. Characterization of miR‐199b expression pattern among normal HSPC populations revealed that miR‐199b is enriched in LT‐HSCsand reduced upon myeloablativestress, suggesting its role in HSCsmaintenance. Indeed, our results reveal that loss of miR‐199b5p results in imbalance between long‐term hematopoieticstem cells (LT‐HSCs), short‐term hematopoieticstem cells (ST‐HSCs) and multipotentprogenitors (MMPs) pool. We found that during homeostasis, miR‐199b‐null HSCshave reduced capacity to maintain quiescent state and exhibit cell‐cycle deregulation. Cell cycle analyses showed that attenuation of miR‐199b controls HSCspool, causing defects in G1‐S transition of cell cycle, without significant changes in apoptosis. This might be due to increased differentiation of LT‐HSCs into MPPs. Indeed, cell differentiation assay in vitro showed that FACS‐sorted LT‐HSCs(LineagenegSca1posc‐Kitpos CD48neg CD150pos) lacking miR‐199b have increased differentiation potential into MPP in the presence of early cytokines. In addition, differentiation assays in vitro in FACS‐sorted LSK population of 52 weeks old miR‐199b KO mice revealed that loss of miR‐199b promotes accumulation of GMP‐like progenitors but decreases lymphoid differentiation, suggesting that miR199b may regulate age‐related pathway. We used noncompetitive repopulation studies to show that overall BM donor cellularitywas markedly elevated in the absence of miR‐199b among HSPCs, committed progenitors and mature myeloid but not lymphoid cell compartments. This may suggest that miR‐199b‐null LT‐HSC render enhanced self‐renewal capacity upon regeneration demand yet promoting myeloid reconstitution. Moreover, when we challenged the self‐renewal potential of miR‐199b‐null LT‐HSC by a secondary BM transplantation of unfractionatedBM cells from primary recipients into secondary hosts, changes in PB reconstitution were dramatic. Gating for HSPCspopulations in the BM of secondary recipients in 24 weeks after BMT revealed that levels of LT‐HSC were similar between recipients reconstituted with wild‐type and miR‐199b‐KO chimeras, whereas miR‐199b‐null HSCscontributed relatively more into MPPs. Our data identify that attenuation of miR‐199b leads to loss of quiescence and premature differentiation of HSCs. These findings indicate that loss of miR‐199b promotes signals that govern differentiation of LT‐HSC to MPP leading to accumulation of highly proliferativeprogenitors during long‐term reconstitution. Hematopoieticregeneration via repopulation studies also revealed that miR‐199b‐deficient HSPCshave a lineage skewing potential toward myeloid lineage or clonalmyeloid bias, a hallmark of aging HSCs, implicating a regulatory role for miR‐199b in hematopoietic aging

Cryptic collagen IV promotes cell migration and adhesion in myeloid leukemia.

Previously, we showed that discoidin domain receptor 1 (DDR1), a class of collagen-activated receptor tyrosine kinase (RTK) was highly upregulated on bone marrow (BM)-derived CD33+ leukemic blasts of acute myeloid leukemia (AML) patients. Herein as DDR1 is a class of collagen-activated RTK, we attempt to understand the role of native and remodeled collagen IV in BM microenvironment and its functional significance in leukemic cells. Exposure to denatured collagen IV significantly increased the migration and adhesion of K562 cells, which also resulted in increased activation of DDR1 and AKT. Further, levels of MMP9 were increased in conditioned media (CM) of denatured collagen IV exposed cells. Mass spectrometric liquid chromatography/tandem mass spectrometry QSTAR proteomic analysis revealed exclusive presence of Secretogranin 3 and InaD-like protein in the denatured collagen IV CM. Importantly, BM samples of AML patients exhibited increased levels of remodeled collagen IV compared to native as analyzed via anti-HUIV26 antibody. Taken together, for the first time, we demonstrate that remodeled collagen IV is a potent activator of DDR1 and AKT that also modulates both migration and adhesion of myeloid leukemia cells. Additionally, high levels of the HUIV26 cryptic collagen IV epitope are expressed in BM of AML patients. Further understanding of this phenomenon may lead to the development of therapeutic agents that directly modulate the BM microenvironment and attenuate leukemogenesis

miR-125a regulates cell cycle, proliferation, and apoptosis by targeting the ErbB pathway in acute myeloid leukemia.

microRNA profiling of acute myeloid leukemia patient samples identified miR-125a as being decreased. Current literature has investigated miR-125a\u27s role in normal hematopoiesis but not within acute myeloid leukemia. Analysis of the upstream region of miR-125a identified several CpG islands. Both precursor and mature miR-125a increased in response to a de-methylating agent, Decitabine. Profiling revealed the ErbB pathway as significantly decreased with ectopic miR-125a. Either ectopic expression of miR-125a or inhibition of ErbB via Mubritinib resulted in inhibition of cell cycle proliferation and progression with enhanced apoptosis revealing ErbB inhibitors as potential novel therapeutic agents for treating miR-125a-low AML