Biological network-driven gene selection identifies a stromal immune module as a key determinant of triple-negative breast carcinoma prognosis.

Triple-negative breast cancer (TNBC) is a heterogeneous group of aggressive breast cancers for which no targeted treatment is available. Robust tools for TNBC classification are required, to improve the prediction of prognosis and to develop novel therapeutic interventions. We analyzed 3,247 primary human breast cancer samples from 21 publicly available datasets, using a five-step method: (1) selection of TNBC samples by bimodal filtering on ER-HER2 and PR, (2) normalization of the selected TNBC samples, (3) selection of the most variant genes, (4) identification of gene clusters and biological gene selection within gene clusters on the basis of String© database connections and gene-expression correlations, (5) summarization of each gene cluster in a metagene. We then assessed the ability of these metagenes to predict prognosis, on an external public dataset (METABRIC). Our analysis of gene expression (GE) in 557 TNBCs from 21 public datasets identified a six-metagene signature (167 genes) in which the metagenes were enriched in different gene ontologies. The gene clusters were named as follows: Immunity1, Immunity2, Proliferation/DNA damage, AR-like, Matrix/Invasion1 and Matrix2 clusters respectively. This signature was particularly robust for the identification of TNBC subtypes across many datasets (n = 1,125 samples), despite technology differences (Affymetrix© A, Plus2 and Illumina©). Weak Immunity two metagene expression was associated with a poor prognosis (disease-specific survival; HR = 2.68 [1.59-4.52], p = 0.0002). The six-metagene signature (167 genes) was validated over 1,125 TNBC samples. The Immunity two metagene had strong prognostic value. These findings open up interesting possibilities for the development of new therapeutic interventions.

Vestibular inputs to propriospinal interneurons in the feline C1-C2 spinal cord projecting to the C5-C6 ventral horn.

The resting length of respiratory muscles must be altered during changes in posture in order to maintain stable ventilation. Prior studies showed that although the vestibular system contributes to these adjustments in respiratory muscle activity, the medullary respiratory groups receive little vestibular input. Additionally, previous transneuronal tracing studies demonstrated that propriospinal interneurons in the C(1)-C(2) spinal cord send projections to the ipsilateral diaphragm motor pool. The present study tested the hypothesis that C(1)-C(2) interneurons mediate vestibular influences on diaphragm activity. Recordings were made from 145 C(1)-C(2) neurons that could be antidromically activated from the ipsilateral C(5)-C(6 )ventral horn, 60 of which had spontaneous activity, during stimulation of vestibular receptors using electric current pulses or whole-body rotations in vertical planes. The firing of 19 of 31 spontaneously active neurons was modulated by vertical vestibular stimulation; the response vector orientations of many of these cells were closer to the pitch plane than the roll plane, and their response gains remained relatively constant across stimulus frequencies. Virtually all spontaneously active neurons responded robustly to electrical vestibular stimulation, and their response latencies were typically shorter than those for diaphragm motoneurons. Nonetheless, respiratory muscle responses to vestibular stimulation were still present after inactivation of the C(1)-C(2) cord using large injections of either muscimol or ibotenic acid. These data suggest that C(1)-C(2) propriospinal interneurons contribute to regulating posturally related responses of the diaphragm, although additional pathways are also involved in generating this activity.

Responses of feline medial medullary reticular formation neurons with projections to the C5-C6 ventral horn to vestibular stimulation.

Prior studies have shown that the vestibular system contributes to adjusting respiratory muscle activity during changes in posture, and have suggested that portions of the medial medullary reticular formation (MRF) participate in generating vestibulo-respiratory responses. However, there was previously no direct evidence to demonstrate that cells in the MRF relay vestibular signals monosynaptically to respiratory motoneurons. The present study tested the hypothesis that the firing of MRF neurons whose axons could be antidromically activated from the vicinity of diaphragm motoneurons was modulated by whole-body rotations in vertical planes that stimulated vestibular receptors, as well as by electrical current pulses delivered to the vestibular nerve. In total, 171 MRF neurons that projected to the C5-C6 ventral horn were studied; they had a conduction velocity of 34+/-15 (standard deviation) m/sec. Most (135/171 or 79%) of these MRF neurons lacked spontaneous firing. Of the subpopulation of units with spontaneous discharges, only 3 of 20 cells responded to vertical rotations up to 10 degrees in amplitude, whereas the activity of 8 of 14 neurons was affected by electrical stimulation of the vestibular nerve. These data support the hypothesis that the MRF participates in generating vestibulo-respiratory responses, but also suggest that some neurons in this region have other functions.