Project description:CD20 protein-negative diseases were observed for a subset of patients with B-NHL that relapsed after initial responses to mosunetuzumab. For many of these tumors, whole exome (WES) and RNA-seq failed to detect any MS4A1 genetic variants or loss of CD20 mRNA, which could have explained the apparent loss of CD20 protein. For this study, we obtained formalin-fixed paraffin-embedded (FFPE) tumor samples from a similar cohort of four additional patients relapsing after mosunetuzumab, and performed RNA-seq to look for changes in the splicing pattern of CD20.
Project description:Resistance to enteric pathogens is a complex trait at the crossroads of multiple biological processes. We have previously shown in the Drosophila Genetic Reference Panel (DGRP) that resistance to infection is highly heritable, but our understanding of how the effects of genetic variants are channeled through distinct molecular layers is still limited. To address this, we employed a systems genetics approach on the gut transcriptomes from 38 control and orally-infected DGRP lines, identifying a large number of condition-specific expression quantitative trait loci (cis-eQTLs). By assessing the allelic imbalance in the transcriptomes of 19 F1 hybrid lines from a large round-robin design, we could independently attribute a robust cis-regulatory effect to only 10% of the detected cis-eQTLs. These results therefore suggest that many context-dependent eQTLs that are assumed to act in cis, may act in trans instead. Further comparison of the transcriptomes of resistant and susceptible DGRP lines revealed Nutcracker (ntc) as the only differentially expressed gene between resistance classes. Interestingly, we found that ntc is linked to infection-specific eQTLs that not only correlate with its expression level, but also to enteric infection susceptibility. This is consistent with our findings that ntc expression is induced upon infection, whereas loss of ntc confers an overall greater susceptibility to infection, including lower innate immune activation as well as impaired infection-induced stem cell activity. Further mechanistic analysis revealed one ntc eQTL that significantly decreases the binding affinity for the repressor Broad, resulting into an allele-specific ntc expression increase. Our collective findings therefore point to a large number of infection-specific cis and trans-acting eQTLs in the DGRP, including one common non-coding variant that lowers enteric infection susceptibility by modulating ntc gene regulation through altered Broad repressor binding.
Project description:To assess a potential role of transcription factor CREM in the long-term detrimental effects of beta1-adrenoceptor overexpression, four mouse lines were generated and studied: wild-type mice (WT), Crem-normal beta1AR-transgenic mice (beta1ARTG), Crem-deficient non-transgenic mice (Crem-/-) and Crem-deficient beta1AR-transgenic mice (beta1ARTG/Crem-/-). We focused on genes up- or down-regulated in transgenic mice due to the lacking of CREM (beta1ARTG/Crem-/- vs. beta1ARTG).
Project description:The mammalian olfactory system detects and discriminates between millions of odorants to elicit appropriate behavioral responses. While much has been learned about how olfactory sensory neurons detect odorants and signal their presence, how specific innate, unlearned behaviors are initiated in response to ethologically relevant odors remains poorly understood. Here, we show that the 4-transmembrane protein CD20, also known as MS4A1, is expressed in a previously uncharacterized subpopulation of olfactory sensory neurons in the main olfactory epithelium of the murine nasal cavity and functions as a mammalian odorant receptor that recognizes compounds produced by mouse predators. While wildtype mice avoid these predator odorants, mice genetically deleted of CD20 do not appropriately respond. Together, this work reveals a novel CD20-mediated odor-sensing mechanism in the mammalian olfactory system that triggers innate behaviors critical for organismal survival.
Project description:For the complete cure of tumors, it is essential to eliminate cancer-initiating cells (CICs). Immunohistochemically, most tumor cells were CD20 and/or CD138 positive in clinical samples of lymphoplasmacytic lymphoma (LPL), and CD20- CD138- cells were hardly detected. Therefore, useful positive markers expressing in a candidate of CICs of LPL are necessary. First, we performed gene expression microarray analysis between CD20- CD138- and CD20+ CD138+ subpopulations using sorted Waldenstrom macroglobulinemia cell line (MWCL-1).
Project description:Water homeostasis is regulated by the peptide hormone arginine vasopressin (AVP), which promotes water reabsorption in the renal collecting duct. The regulation of Aqp2 gene transcription is a key mechanism through which AVP modulates water transport as disruption of this mechanism leads to water balance disorders. Therefore, an important goal is to understand the regulatory processes that control Aqp2 gene transcription. While CREB (CREB1) has been proposed as the primary transcription factor responsible for Aqp2 transcription, recent evidence challenges this view, suggesting that other CREB-like transcription factors, including ATF1 and CREM, may play a role. We employed the CRISPR/Cas9 gene-editing system to delete Atf1, Creb1, and Crem in mpkCCD cells, an immortalized collecting duct cell line. These cell lines were then exposed to the vasopressin analog dDAVP to assess the role of these transcription factors in regulating Aqp2 expression. Deletion of all three transcription factors (ATF1, CREB1, and CREM) led to a significant reduction in the vasopressin-induced upregulation of AQP2 protein, confirming their role in regulating Aqp2 expression. Rescue experiments in triple knockout cells showed that expressing any of the three transcription factors restored the response to vasopressin. RNA-seq data showed that Aqp2 mRNA levels mirrored changes in protein abundance, supporting the idea that these transcription factors affect Aqp2 transcription. Our findings demonstrate that ATF1, CREB1, and CREM have redundant roles in regulating Aqp2 transcription. Our results suggest that these transcription factors might regulate the expression of other unidentified transcription factors involved in Aqp2 regulation.
