Deregulation of MYCN, LIN28B and LET7 in a Molecular Subtype of Aggressive High-Grade Serous Ovarian Cancers

Molecular subtypes of serous ovarian cancer have been recently described. Using data from independent datasets including over 900 primary tumour samples, we show that deregulation of the Let-7 pathway is specifically associated with the C5 molecular subtype of serous ovarian cancer. DNA copy number and gene expression of HMGA2, alleles of Let-7, LIN28, LIN28B, MYC, MYCN, DICER1, and RNASEN were measured using microarray and quantitative reverse transcriptase PCR. Immunohistochemistry was performed on 127 samples using tissue microarrays and anti-HMGA2 antibodies. Fluorescence in situ hybridisation of bacterial artificial chromosomes hybridized to 239 ovarian tumours was used to measure translocation at the LIN28B locus. Short interfering RNA knockdown in ovarian cell lines was used to test the functionality of associations observed. Four molecular subtypes (C1, C2, C4, C5) of high-grade serous ovarian cancers were robustly represented in each dataset and showed similar pattern of patient survival. We found highly specific activation of a pathway involving MYCN, LIN28B, Let-7 and HMGA2 in the C5 molecular subtype defined by MYCN amplification and over-expression, over-expression of MYCN targets including the Let-7 repressor LIN28B, loss of Let-7 expression and HMGA2 amplification and over-expression. DICER1, a known Let-7 target, and RNASEN were over-expressed in C5 tumours. We saw no evidence of translocation at the LIN28B locus in C5 tumours. The reported interaction between LIN28B and Let-7 was recapitulated by siRNA knockdown in ovarian cancer cell lines. Our results associate deregulation of MYCN and downstream targets, including Let-7 and oncofetal genes, with serous ovarian cancer. We define for the first time how elements of an oncogenic pathway, involving multiple genes that contribute to stem cell renewal, is specifically altered in a molecular subtype of serous ovarian cancer. By defining the drivers of a molecular subtype of serous ovarian cancers we provide a novel strategy for targeted therapeutic intervention

Intravenous injection of neural progenitor cells improved depression-like behavior after cerebral ischemia

Poststroke depression (PSD) occurs in approximately one-third of stroke survivors and is one of the serious sequelae of stroke. The onset of PSD causes delayed functional recovery by rehabilitation and also increases cognitive impairment. However, appropriate strategies for the therapy against ischemia-induced depression-like behaviors still remain to be developed. Such behaviors have been associated with a reduced level of brain-derived neurotrophic factor (BDNF). In addition, accumulating evidence indicates the ability of stem cells to improve cerebral ischemia-induced brain injuries. However, it remains to be clarified as to the effect of neural progenitor cells (NPCs) on PSD and the association between BDNF level and PSD. Using NPCs, we investigated the effect of intravenous injection of NPCs on PSD. We showed that injection of NPCs improved ischemia-induced depression-like behaviors in the forced-swimming test and sucrose preference test without having any effect on the viable area between vehicle- and NPC-injected ischemic rats. The injection of NPCs prevented the decrease in the level of BDNF in the ipsilateral hemisphere. The levels of phosphorylated CREB, ERK and Akt, which have been implicated in events downstream of BDNF signaling, were also decreased after cerebral ischemia. NPC injection inhibited these decreases in the phosphorylation of CREB and ERK, but not that of Akt. Our findings provide evidence that injection of NPCs may have therapeutic potential for the improvement of depression-like behaviors after cerebral ischemia and that these effects might be associated with restoring BDNF-ERK-CREB signaling

Transitional Flow in a Cylindrical Flow Chamber for Studies at the Cellular Level

Fluid shear stress is an important regulator of vascular and endothelial cell (EC) functions. Its effect is dependent not only on magnitude but also on flow type. Although laminar flow predominates in the vasculature, transitional flow can occur and is thought to play a role in vascular diseases. While a great deal is known about the mechanisms and signaling cascades through which laminar shear stress regulates cells, little is known on how transitional shear stress regulates cells. To better understand the response of endothelial cells to transitional shear stress, a novel cylindrical flow chamber was designed to expose endothelial cells to a transitional flow environment similar to that found in vivo. The velocity profiles within the transitional flow chamber at Reynolds numbers 2200 and 3000 were measured using laser Doppler anemometry (LDA). At both Reynolds numbers, the velocity profiles are blunt (non-parabolic) with fluctuations larger than 5% of the velocity at the center of the pipe indicating the flows are transitional. Based on near wall velocity measurements and well established data for flow at these Reynolds numbers, the wall shear stress was estimated to be 3–4 and 5–6 dynes/cm(2) for Reynolds number 2200 and 3000, respectively. In contrast to laminar shear stress, no cell alignment was observed under transitional shear stress at both Reynolds numbers. However, transitional shear stress at the higher Reynolds number caused cell elongation similar to that of laminar shear stress at 3 dynes/cm(2). The fluctuating component of the wall shear stress may be responsible for these differences. The transitional flow chamber will facilitate cellular studies to identify the mechanisms through which transitional shear stress alters EC biology, which will assist in the development of vascular therapeutic treatments