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Biology subjects

Alfonso, A.

Publications and source records attributed to Alfonso, A..

3 recordsLinked to original sources

HLH-3 Is Required for the Maturation of Neurons Necessary for Male Specific Exploration Behavior

Sexually dimorphic gene expression within the nervous system of many organisms gives rise to features that underline sexually dimorphic behaviors. In C. elegans, terminal differentiation of neurons with sexually dimorphic features often occur not at the time of neuronal birth, but rather during the juvenile to adult stage transition, when sexual maturation occurs. Previous works have shown that the bHLH transcription factor, HLH-3, is necessary for the differentiation of sex-specific neurons in hermaphrodites. Expression of the bHLH transcription factor, hlh-3, is dynamic and widespread throughout the embryo. Post-embryonic expression in the hermaphrodite is restricted to the HSNs, and the P cells and their descendants, including the VCs. In this work, we characterize the post-embryonic expression pattern of hlh-3 in the male. Our findings show that the pattern of expression observed in males varies dramatically from that observed in hermaphrodites. Males show expanded expression in the head at the L1 and late L3/early L4 stage, as well as broader expression in the tail at the early L3/Late L4 stage. This leads us to the conclusion that the post-embryonic expression pattern of hlh-3 is sexually dimorphic. We also describe the role of hlh-3 in the maturation of a subset of neurons required for male specific exploration behavior. Our findings show that in the absence of hlh-3 function, the AWA, ASJ, and AIM neurons fail to acquire terminally differentiated features and therefore contribute to the failure of males to perform behaviors mediated by these cells.

genetics↗

Phenotyping the tumor-immune microenvironment (TiME) in vivo by cellular-level optical imaging reveals unique combination phenotypes with variable inflammation and endothelial anergy

Immunotherapies have shown unprecedented clinical benefits in several malignancies1-3. However, clinical responses remain variable and unpredictable, indicating the need to develop predictive platforms that can improve patient stratification4. Phenotyping of tumors into hot, altered, or cold5 based on assessment of only T-cell infiltration in static tumor biopsies provides suboptimal prediction of immunotherapy response6,7. In vivo dynamic mechanisms within the tumor microenvironment such as tumor angiogenesis and leukocyte trafficking5,8,9 also play a central role in modulating anti-tumor immunity and therefore immunotherapy response. Here, we report novel tumor immune microenvironment (TiME) phenotyping in vivo in patients with non-invasive spatially-resolved cellular-level imaging based on endogenous contrast. Investigating skin cancers as a model, with reflectance confocal microscopy (RCM) imaging10, we determined four major phenotypes with variable prevalence of vasculature (Vasc) and inflammation (Inf) features: VaschiInfhi, VaschiInflo, VascloInfhi and Vascmed/hiInflo. The VaschiInfhi phenotype correlates with high immune activation, exhaustion, and vascular signatures while VaschiInflo with endothelial anergy and immune exclusion. Automated quantification of TiME features demonstrates moderate-high accuracy and correlation with corresponding gene expression. Prospectively analyzed response to topical immunotherapy show highest response in VascloInfhi, and reveals the added value of vascular features in predicting treatment response. Our novel in vivo cellular-level imaging and phenotyping approach can potentially advance our fundamental understanding of TiME, develop robust predictors for immunotherapy outcomes and identify novel targetable pathways in future.

cancer biology↗

The conserved ASCL1/MASH-1 ortholog HLH-3 specifies sex-specific ventral cord motor neuron fate in C. elegans

Neural specification can be regulated by one or many transcription factors. Here we identify a novel role for one conserved proneural factor, the bHLH protein HLH-3, implicated in the specification of sex-specific ventral cord motor neurons in C. elegans. In the process of characterizing the role of hlh-3 in neural specification, we document that differentiation of the ventral cord type C neurons, VCs, within their motor neuron class, is dynamic in time and space. Expression of VC class-specific and subclass-specific identity genes is distinct through development and dependent on where they are along the A-P axis (and their position in proximity to the vulva). Our characterization of the expression of VC class and VC subclass-specific differentiation markers in the absence of hlh-3 function reveals that VC fate specification, differentiation, and morphology requires hlh-3 function. Finally, we conclude that hlh-3 cell-autonomously specifies VC cell fate.

genetics↗