miR-133a-A Potential Target for Improving Cardiac Mitochondrial Health and Regeneration After Injury.

ABSTRACT: The various roles of muscle secretory factors and myokines have been well studied, but in recent decades, the role of myocyte-specific microRNAs (myomiRs) has gained momentum. These myomiRs are known to play regulatory roles in muscle health in general, both skeletal muscle and cardiac muscle. In this review, we have focused on the significance of a myomiR termed miR-133a in cardiovascular health. The available literature supports the claim that miR-133a could be helpful in the healing process of muscle tissue after injury. The protective function could be due to its regulatory effect on muscle or stem cell mitochondrial function. In this review, we have shed light on the protective mechanisms offered by miR-133a. Most of the beneficial effects are due to the presence of miR-133a in circulation or tissue-specific expression. We have also reviewed the potential mechanisms by which miR-133a could interact with cell surface receptors and also transcriptional mechanisms by which they offer cardioprotection and regeneration. Understanding these mechanisms will help in finding an ideal strategy to repair cardiac tissue after injury.

HOXA/PBX3 knockdown impairs growth and sensitizes cytogenetically normal acute myeloid leukemia cells to chemotherapy.

The cytogenetically normal subtype of acute myeloid leukemia is associated with an intermediate risk which complicates therapeutic options. Lower overall HOX/TALE expression appears to correlate with more favorable prognosis/better response to treatment in some leukemias and solid cancer. The functional significance of the associated gene expression and response to chemotherapy is not known. Three independent microarray datasets obtained from large cohorts of patients along with quantitative polymerase chain reaction validation were used to identify a four-gene HOXA/TALE signature capable of prognostic stratification. Biochemical analysis was used to identify interactions between the four encoded proteins and targeted knockdown used to examine the functional importance of sustained expression of the signature in leukemia maintenance and response to chemotherapy. An 11 HOXA/TALE code identified in an intermediate-risk group of patients (n=315) compared to a group with a favorable risk (n=105) was reduced to a four-gene signature of HOXA6, HOXA9, PBX3 and MEIS1 by iterative analysis of independent platforms. This signature maintained the favorable/intermediate risk partition and where applicable, correlated with overall survival in cytogenetically normal acute myeloid leukemia. We further showed that cell growth and function are dependent on maintained levels of these core genes and that direct targeting of HOXA/PBX3 sensitizes cytogenetically normal acute myeloid leukemia cells to standard chemotherapy. Together the data support a key role for HOXA/TALE in cytogenetically normal acute myeloid leukemia and demonstrate that targeting of clinically significant HOXA/PBX3 elements may provide therapeutic benefit to patients with this subtype of leukemia.

Elevated TRIB2 with NOTCH1 activation in paediatric/adult T-ALL.

TRIB2 is a potent oncogene, elevated in a subset of human acute myeloid leukaemias (AML) with a mixed myeloid/lymphoid phenotype and NOTCH1 mutations. Although rare in AML, activating NOTCH1 mutations occur in 50% of all T cell acute lymphoblastic leukaemias (T-ALL). TRIB2 is a NOTCH1 target gene that functions in the degradation of key proteins and modulation of MAPK signalling pathways, implicated in haematopoietic cell survival and proliferation. This study showed that TRIB2 expression level is highest in the lymphoid compartment of normal haematopoietic cells, specifically in T cells. Analysis of TRIB2 expression across 16 different subtypes of human leukaemia demonstrated that TRIB2 expression was higher in ALL phenotypes versus all other phenotypes including AML, chronic lymphocytic leukaemia (CLL), myelodysplastic syndrome (MDS) and chronic myeloid leukaemia (CML). A T cell profile was distinguished by high TRIB2 expression in normal and malignant haematopoiesis. High TRIB2 expression was seen in T-ALL with normal karyotype and correlated with NOTCH signalling pathways. High TRIB2 expression correlated with NOTCH1/FBXW7 mutations in a paediatric T-ALL cohort, strongly linking NOTCH1 activation and high TRIB2 expression in paediatric T-ALL. The relationship between TRIB2 and T cell signalling pathways uniquely identifies leukaemia subtypes and will be useful in the advancement of our understanding of T cell and ALL biology.

PGC-1alpha is coupled to HIF-1alpha-dependent gene expression by increasing mitochondrial oxygen consumption in skeletal muscle cells.

Mitochondrial biogenesis occurs in response to increased cellular ATP demand. The mitochondrial electron transport chain requires molecular oxygen to produce ATP. Thus, increased ATP generation after mitochondrial biogenesis results in increased oxygen demand that must be matched by a corresponding increase in oxygen supply. We found that overexpression of peroxisome proliferator-activated receptor-gamma coactivator 1alpha (PGC-1alpha), which increases mitochondrial biogenesis in primary skeletal muscle cells, leads to increased expression of a cohort of genes known to be regulated by the dimeric hypoxia-inducible factor (HIF), a master regulator of the adaptive response to hypoxia. PGC-1alpha-dependent induction of HIF target genes under physiologic oxygen concentrations is not through transcriptional coactivation of HIF or up-regulation of HIF-1alpha mRNA but through HIF-1alpha protein stabilization. It occurs because of intracellular hypoxia as a result of increased oxygen consumption after mitochondrial biogenesis. Thus, we propose that at physiologic oxygen concentrations, PGC-1alpha is coupled to HIF signaling through the regulation of intracellular oxygen availability, allowing cells and tissues to match increased oxygen demand after mitochondrial biogenesis with increased oxygen supply.

Effect of gestational stage on uterine artery blood flow during exercise in rabbits.

We tested the hypothesis that the uterine artery vasoconstrictor response to graded exercise during early gestation would be similar to the nonpregnant (NP) state and would be attenuated at mid and term gestation. Responses to graded treadmill exercise were measured in six female New Zealand White rabbits in the NP state and at day 10, day 20, and day 28 (term) of gestation. Uterine artery blood flow (UtBF) was measured continuously with a Transonic flow probe. Rabbits performed a graded exercise test to voluntary exhaustion (maximal exercise) starting at 7 m/min, 7% grade. UtBF and uterine artery conductance (UtC) decreased similarly during graded exercise (P < 0.01) in the NP state [at maximal exercise: -40% (SD 20) for UtBF and -45% (SD 14) for UtC] and at day 10 of gestation [at maximal exercise: -48% (SD 17) for UtBF; -56% (SD 14) for UtC]. In contrast, there was little change in UtBF or UtC during graded exercise at day 20 [at maximal exercise: -4% (SD 17) for UtBF, P < 0.05 vs. NP; -16% (SD 12) for UtC, P < 0.01 vs. NP] and at day 28 [At maximal exercise: +7% (SD 15) for UtBF, P < 0.01 vs. NP; -2% (SD 24) for UtC, P < 0.01 vs. NP], indicating substantial attenuation of the uterine artery vasoconstrictor response to exercise. At rest, UtC responses to graded doses of intravenous phenylephrine were unaffected by the stage of gestation, which suggests that uterine artery responsiveness to alpha(1)-adrenoreceptor stimulation is preserved through gestation. Normal pregnancy in the rabbit is associated with attenuation of the uterine artery vasoconstrictor response to graded exercise that develops by mid gestation.

Effect of pregnancy on the uterine vasoconstrictor response to exercise in rats.

A major maternal adaptation in pregnancy is the large increase in uteroplacental blood flow that supplies the growing fetus with oxygen and nutrients. The impact of gestation on the dynamic uterine vasoconstrictor response to exercise in the rat, a common model for pathophysiological disorders in pregnancy remains unknown. We hypothesized that rats exhibit a robust uterine vasoconstrictor response to acute exercise that is attenuated in late pregnancy. Pregnant (P, N = 12) and nonpregnant (NP, N = 8) rats were instrumented chronically with a ultrasonic transit-time flowprobe and carotid arterial catheter to directly measure uterine artery blood flow (UtBF) and blood pressure (BP), respectively, at day 20 of gestation for 5 min of treadmill exercise (7 m/min; 6% grade). Preexercise UtBF [P, 2.1 (SD1.6) vs. NP, 0.5 (SD0.3) mL/min P < 0.01) and uterine artery conductance (UtC) [P, 2.1(SD1.7) vs. NP, 0.4 (SD0.2) mL/min × mmHg(-1) × 10(-2), P < 0.01] were higher in pregnant rats, whereas preexercise BP was lower in the pregnant rats [P, 111 (SD13) vs. NP, 126 (SD13) mmHg, P = 0.02]. Preexercise heart rate was similar [P, 457 (SD30) vs. NP, 454 (SD42), P = 0.3]. Exercise initiated rapid and sustained decreases in UtBF [Δ-47% (SD12)] and UtC [Δ-49% (SD12)] that were attenuated in the pregnant rats [UtBF, Δ-25% (SD20) and UtC, Δ-30% (SD20), P = 0.02]. The BP and heart rate responses to exercise were unaffected in late pregnancy (interaction term, P = 0.3). In rats, dynamic exercise induces a uterine vasoconstrictor response that is blunted during late gestation, a response that we observed previously in pregnant rabbits.

Potential prognostic marker ubiquitin carboxyl-terminal hydrolase-L1 does not predict patient survival in non-small cell lung carcinoma.

BACKGROUND: Ubiquitin Carboxyl-Terminal Hydrolase-L1 (UCH-L1) is a deubiquitinating enzyme that is highly expressed throughout the central and peripheral nervous system and in cells of the diffuse neuroendocrine system. Aberrant function of UCH-L1 has been associated with neurological disorders such as Parkinson's disease and Alzheimer's disease. Moreover, UCH-L1 exhibits a variable expression pattern in cancer, acting either as a tumour suppressor or promoter, depending on the type of cancer. In non-small cell lung carcinoma primary tumour samples, UCH-L1 is highly expressed and is associated with an advanced tumour stage. This suggests UCH-L1 may be involved in oncogenic transformation and tumour invasion in NSCLC. However, the functional significance of UCH-L1 in the progression of NSCLC is unclear. The aim of this study was to investigate the role of UCH-L1 using NSCLC cell line models and to determine if it is clinically relevant as a prognostic marker for advanced stage disease. METHODS: UCH-L1 expression in NSCLC cell lines H838 and H157 was modulated by siRNA-knockdown, and the phenotypic changes were assessed by flow cytometry, haematoxylin & eosin (H&E) staining and poly (ADP-ribose) polymerase (PARP) cleavage. Metastatic potential was measured by the presence of phosphorylated myosin light chain (MLC2). Tumour microarrays were examined immunohistochemically for UCH-L1 expression. Kaplan-Meier curves were generated using UCH-L1 expression levels and patient survival data extracted from Gene Expression Omnibus data files. RESULTS: Expression of UCH-L1 was decreased by siRNA in both cell lines, resulting in increased cell death in H838 adenocarcinoma cells but not in the H157 squamous cell line. However, metastatic potential was reduced in H157 cells. Immunohistochemical staining of UCH-L1 in patient tumours confirmed it was preferentially expressed in squamous cell carcinoma rather than adenocarcinoma. However the Kaplan-Meier curves generated showed no correlation between UCH-L1 expression levels and patient outcome. CONCLUSIONS: Although UCH-L1 appears to be involved in carcinogenic processes in NSCLC cell lines, the absence of correlation with patient survival indicates that caution is required in the use of UCH-L1 as a potential prognostic marker for advanced stage and metastasis in lung carcinoma.

Identification of gene networks associated with acute myeloid leukemia by comparative molecular methylation and expression profiling.

Around 80% of acute myeloid leukemia (AML) patients achieve a complete remission, however many will relapse and ultimately die of their disease. The association between karyotype and prognosis has been studied extensively and identified patient cohorts as having favourable [e.g. t(8; 21), inv (16)/t(16; 16), t(15; 17)], intermediate [e.g. cytogenetically normal (NK-AML)] or adverse risk [e.g. complex karyotypes]. Previous studies have shown that gene expression profiling signatures can classify the sub-types of AML, although few reports have shown a similar feature by using methylation markers. The global methylation patterns in 19 diagnostic AML samples were investigated using the Methylated CpG Island Amplification Microarray (MCAM) method and CpG island microarrays containing 12,000 CpG sites. The first analysis, comparing favourable and intermediate cytogenetic risk groups, revealed significantly differentially methylated CpG sites (594 CpG islands) between the two subgroups. Mutations in the NPM1 gene occur at a high frequency (40%) within the NK-AML subgroup and are associated with a more favourable prognosis in these patients. A second analysis comparing the NPM1 mutant and wild-type research study subjects again identified distinct methylation profiles between these two subgroups. Network and pathway analysis revealed possible molecular mechanisms associated with the different risk and/or mutation sub-groups. This may result in a better classification of the risk groups, improved monitoring targets, or the identification of novel molecular therapies.

Uterine artery blood flow and renal sympathetic nerve activity during exercise in rabbit pregnancy.

The uterine artery blood flow (UtBF) and renal sympathetic nerve activity (SNA) responses to treadmill exercise were evaluated in 12 nonpregnant (NP) and 17 term pregnant (P) rabbits. UtBF was monitored continuously with a Transonic flowprobe. Rabbits underwent three exercise trials (5-min duration) that varied in absolute workload. The rise in renal SNA with exercise was intensity related. Pregnancy did not affect the average steady-state renal SNA response expressed relative to maximum activity (P 24 +/- 1% vs. NP 23 +/- 2% of maximum smoke-elicited activity) and increased the average renal SNA response expressed relative to resting activity (P +155 +/- 19% vs. NP +84 +/- 23% from rest, P = 0.03) At rest, UtBF (P 13 +/- 3 vs. NP 1.9 +/- 0.3 ml/min) and uterine artery conductance (UtC; P 22 +/- 5 vs. NP 2.8 +/- 0.5 ml. min-1.mmHg-1 x 10-2) were elevated in the P rabbits. The average exercise-related decreases in UtBF (P -16 +/- 4% vs. NP -48 +/- 4%) and UtC (P -27 +/- 4% vs. NP -54 +/- 4%) were attenuated in the P rabbits. Pregnancy does not impair the ability to raise renal SNA but attenuates the uterine artery constrictor response to moderate to heavy dynamic exercise in rabbits. Under normal conditions, the pregnant uterine circulatory bed may be relatively protected from exercise-related redistribution of blood flow.

Pregnancy and acute baroreflex resetting in conscious rabbits.

To test the hypothesis that acute resetting of baroreflex control of heart rate (HR) is enhanced during pregnancy, we determined whether the rightward shift in the baroreflex relationship between arterial pressure and HR after arterial pressure is raised [~25 mmHg for 30 min, due to infusion of phenylephrine (PE) or methoxamine (Meth)] is greater in late pregnant compared with nonpregnant conscious rabbits. Baroreflex function was assessed by monitoring HR responses to both stepwise steady-state changes (n = 14) and rapid ramp changes (n = 10) in arterial pressure. Pregnancy decreased reflex gain, increased reflex minimum HR, and shifted the curves to a lower pressure level, when either the steady-state or ramp method was used (all changes, P < 0.05). When PE was used to increase pressure, resetting of steady-state curves was observed both before and during pregnancy, but the magnitude of the resetting was less in the pregnant rabbits. Further inspection of the data revealed that the size of the shift in pregnant rabbits was inversely related to the dose of PE. Because the pressure rise was the same in all experiments, PE appears to nonspecifically counteract acute resetting. When Meth was used instead to increase pressure, resetting of steady-state curves was similar in pregnant and nonpregnant rabbits and was unrelated to dose. Similarly, when reflex curves were generated using the ramp method, and either Meth or low doses of PE were used to increase pressure, no differences in the degree of resetting were observed between pregnant and nonpregnant rabbits. In summary, high doses of PE counteract acute resetting of baroreflex control of HR. More importantly, while baroreflex function is depressed, the ability of the baroreflex to reset appears to be preserved during pregnancy.