Comprehensive Search for Novel Circulating miRNAs and Axon Guidance Pathway Proteins Associated with Risk of End Stage Kidney Disease in Diabetes

Journal of the American Society of Nephrology : JASN

2021 Jun 17

Satake, E;Saulnier, PJ;Kobayashi, H;Gupta, M;Looker, H;Wilson, J;Md Dom, Z;Ihara, K;O'Neil, K;Krolewski, B;Pipino, C;Pavkov, M;Nair, V;Bitzer, M;Niewczas, M;Kretzler, M;Mauer, M;Doria, A;Najafian, B;Kulkarni, R;Duffin, K;Pezzolesi, M;Kahn, CR;Nelson, R;Krolewski, A;
PMID: 34140396 | DOI: 10.1681/ASN.2021010105

Background Mechanisms underlying the progression of diabetic kidney disease to end-stage kidney disease (ESKD) are not fully understood. Methods We performed global micro-RNA (miRNA) analysis in plasma in two cohorts encompassing 375 individuals with type 1 and type 2 diabetes with late diabetic kidney disease and targeted proteomics analysis in plasma in four cohorts encompassing 746 individuals with late and early diabetic kidney disease. We examined structural lesions in kidney biopsies from the 105 individuals with early diabetic kidney disease. Human umbilical vein endothelial cells were used to assess the effects of miRNA mimics or inhibitors on regulation of candidate proteins. Results In the late diabetic kidney disease cohorts, we identified 17 circulating miRNAs represented by four exemplars (miR-1287-5p, miR-197-5p, miR-339-5p, miR-328-3p), which were strongly associated with 10-year risk of ESKD. These miRNAs targeted proteins in the axon guidance pathway. Circulating levels of six of these proteins-most notably EFNA4 and EPHA2-were strongly associated with 10-year risk of ESKD in all cohorts. Furthermore, circulating levels of these proteins correlated with severity of structural lesions in kidney biopsies. In contrast, expression levels of genes encoding these proteins had no apparent effects on the lesions. In in vitro experiments, mimics of miR-1287-5p and miR-197-5p and inhibitors of miR-339-5p and miR328-3p upregulated concentrations of EPHA2 in either cell lysate, supernatant, or both. Conclusions This study reveals novel mechanisms involved in progression to ESKD and points to the importance of systemic factors in the development of diabetic kidney disease. Some circulating miRNAs and axon guidance pathway proteins represent potential targets for new therapies to prevent and treat this condition.

HIV DNA reservoir and elevated PD-1 expression of CD4 T-cell subsets particularly persist in the terminal ileum of HIV-positive patients despite cART

HIV medicine

2021 Jan 09

Horn, C;Augustin, M;Ercanoglu, MS;Heger, E;Knops, E;Bondet, V;Duffy, D;Chon, SH;Nierhoff, D;Oette, M;Schäfer, H;Vivaldi, C;Held, K;Anderson, J;Geldmacher, C;Suárez, I;Rybniker, J;Klein, F;Fätkenheuer, G;Müller-Trutwin, M;Lehmann, C;
PMID: 33421299 | DOI: 10.1111/hiv.13031

Despite its importance as an HIV anatomic sanctuary, little is known about the characteristics of the HIV reservoir in the terminal ileum (TI). In blood, the immune checkpoint inhibitor programmed-death-1 (PD-1) has been linked to the HIV reservoir and T-cell immune dysfunction. We thus evaluated PD-1 expression and cell-associated HIV DNA in memory CD4 T-cell subsets from TI, peripheral blood (PB) and rectum (RE) of untreated and treated HIV-positive patients to identify associations between PD-1 and HIV reservoir in other sites. Using mononuclear cells from PB, TI and RE of untreated HIV-positive (N = 6), treated (n = 18) HIV-positive and uninfected individuals (n = 16), we identified and sorted distinct memory CD4 T-cell subsets by flow cytometry, quantified their cell-associated HIV DNA using quantitative PCR and assessed PD-1 expression levels using geometric mean fluorescence intensity. Combined HIV-1 RNA in situ hybridization and immunohistochemistry was performed on ileal biopsy sections. Combined antiretroviral therapy (cART)-treated patients with undetectable HIV RNA and significantly lower levels of HIV DNA in PB showed particularly high PD-1 expression in PB and TI, and high HIV DNA levels in TI, irrespective of clinical characteristics. By contrast, in treatment-naïve patients HIV DNA levels in memory CD4 T-cell subsets were high in PB and TI. Elevated PD-1 expression on memory CD4 T-cells in PB and TI despite treatment points to continuous immune dysfunction and underlines the importance of evaluating immunotherapy in reversing HIV latency and T-cell reconstitution. As HIV DNA particularly persists in TI despite cART, investigating samples from TI is crucial in understanding HIV immunopathogenesis.

AIM2 Suppresses Inflammation and Epithelial Cell Proliferation during Glomerulonephritis

Journal of immunology (Baltimore, Md. : 1950)

2021 Dec 01

Chung, H;Komada, T;Lau, A;Chappellaz, M;Platnich, JM;de Koning, HD;Petri, B;Luque, Y;Walker, S;Benediktsson, H;Mesnard, L;Chun, J;Muruve, DA;
PMID: 34740957 | DOI: 10.4049/jimmunol.2100483

Absent in melanoma-2 (AIM2) is an inflammasome-forming innate immune sensor for dsDNA but also exhibits inflammasome-independent functions such as restricting cellular proliferation. AIM2 is expressed in the kidney, but its localization and function are not fully characterized. In normal human glomeruli, AIM2 localized to podocytes. In patients with glomerulonephritis, AIM2 expression increased in CD44+-activated parietal epithelial cells within glomerular crescents. To explore AIM2 effects in glomerular disease, studies in Aim2 -/- mice were performed. Aim2-/- glomeruli showed reduced expression of Wilm tumor gene-1 (WT1), WT1-driven podocyte genes, and increased proliferation in outgrowth assays. In a nephrotoxic serum (NTS)-induced glomerulonephritis model, Aim2-/- (B6) mice exhibited more severe glomerular crescent formation, tubular injury, inflammation, and proteinuria compared with wild-type controls. Inflammasome activation markers were absent in both Aim2 -/- and wild-type kidneys, despite an increased inflammatory transcriptomic signature in Aim2 -/- mice. Aim2 -/- mice also demonstrated dysregulated cellular proliferation and an increase in CD44+ parietal epithelial cells during glomerulonephritis. The augmented inflammation and epithelial cell proliferation in Aim2 -/- (B6) mice was not due to genetic background, as Aim2 -/- (B6.129) mice demonstrated a similar phenotype during NTS glomerulonephritis. The AIM2-like receptor (ALR) locus was necessary for the inflammatory glomerulonephritis phenotype observed in Aim2 -/- mice, as NTS-treated ALR -/- mice displayed equal levels of injury as wild-type controls. Podocyte outgrowth from ALR -/- glomeruli was still increased, however, confirming that the ALR locus is dispensable for AIM2 effects on epithelial cell proliferation. These results identify a noncanonical role for AIM2 in suppressing inflammation and epithelial cell proliferation during glomerulonephritis.

Pathological diagnosis of Coronavirus-related nephropathy: insight from postmortem studies

Critical reviews in clinical laboratory sciences

2021 Jul 08

Sanguedolce, F;Zanelli, M;Froio, E;Bisagni, A;Zizzo, M;Ascani, S;Stallone, G;Netti, S;Ranieri, E;Falagario, U;Carrieri, G;Cormio, L;
PMID: 34236278 | DOI: 10.1080/10408363.2021.1944047

A novel coronavirus pneumonia first occurred in Wuhan, China in early December 2019; the causative agent was identified and named as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by the World Health Organization (WHO), and the resulting disease termed coronavirus disease 2019 (COVID-19), according to the WHO coronavirus disease situation reports. This condition has spread rapidly all over the world and caused more than 125 million cases globally, with more than 2 million related deaths. Two previous outbreaks due to zoonotic coronaviruses have occurred in the last 20 years, namely the severe acute respiratory syndrome coronavirus (SARS-CoV) and the Middle East respiratory syndrome coronavirus (MERS-CoV), causing high morbidity and mortality in human populations upon crossing the species barriers. SARS-CoV-2, SARS-CoV, and MERS-CoV show several similarities in pathogenicity and clinical presentations, the latter ranging from asymptomatic infection to severe acute respiratory distress syndrome (ARDS) and multiorgan impairment. Acute kidney injury (AKI) has been commonly reported in patients with CoV infections; therefore, pathological analysis of renal parenchyma in these patients has been carried out in order to improve knowledge about underlying mechanisms. Viral infection has been demonstrated in the renal tubular epithelial cells by electron microscopy (EM), immunohistochemistry (IHC), and in situ hybridization (ISH), although with conflicting results. Light microscopy (LM) changes have been described in the renal parenchyma primarily in the form of acute renal tubular damage, possibly due to direct viral cytopathic effect and immune-mediated mechanisms such as cytokine storm syndrome. In this review, we describe and discuss the spectrum of histological, ultrastructural, and molecular findings in SARS-CoV, MERS-CoV, and SARS-CoV-2-related renal pathology obtained from postmortem studies, as well as intrinsic limitations and pitfalls of current diagnostic techniques.

Monocyte and macrophage derived myofibroblasts: Is it fate? A review of the current evidence

Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society

2021 Jul 01

Vierhout, M;Ayoub, A;Naiel, S;Yazdanshenas, P;Revill, SD;Reihani, A;Dvorkin-Gheva, A;Shi, W;Ask, K;
PMID: 34107123 | DOI: 10.1111/wrr.12946

Since the discovery of the myofibroblast over 50 years ago, much has been learned about its role in wound healing and fibrosis. Its origin, however, remains controversial, with a number of progenitor cells being proposed. Macrophage-myofibroblast transition (MMT) is a recent term coined in 2014 that describes the mechanism through which macrophages, derived from circulating monocytes originating in the bone marrow, transformed into myofibroblasts and contributed to kidney fibrosis. Over the past years, several studies have confirmed the existence of MMT in various systems, suggesting that MMT could potentially occur in all fibrotic conditions and constitute a reasonable therapeutic target to prevent progressive fibrotic disease. In this perspective, we examined recent evidence supporting the notion of MMT in both human disease and experimental models across organ systems. Mechanistic insight from these studies and information from in vitro studies is provided. The findings substantiating plausible MMT showcased the co-expression of macrophage and myofibroblast markers, including CD68 or F4/80 (macrophage) and α-SMA (myofibroblast), in fibroblast-like cells. Furthermore, fate-mapping experiments in murine models exhibiting myeloid-derived myofibroblasts in the tissue further provide direct evidence for MMT. Additionally, we provide some evidence from single cell RNA sequencing experiments confirmed by fluorescent in situ hybridisation studies, showing monocyte/macrophage and myofibroblast markers co-expressed in lung tissue from patients with fibrotic lung disease. In conclusion, MMT is likely a significant contributor to myofibroblast formation in wound healing and fibrotic disease across organ systems. Circulating precursors including monocytes and the molecular mechanisms governing MMT could constitute valid targets and provide insight for the development of novel antifibrotic therapies; however, further understanding of these processes is warranted.

Association of Matrix Metalloproteinase-2 mRNA Expression with Subtypes of Pediatric Cholesteatoma

BioMed research international

2021 Mar 10

Kan, T;Ueda, H;Takahara, T;Tsuchiya, Y;Kishimoto, M;Uchida, Y;Ogawa, T;Ohashi, W;Tsuzuki, T;Fujimoto, Y;
PMID: 33778077 | DOI: 10.1155/2021/6644897

Cholesteatoma is a clinically heterogeneous disease, with some patients showing spontaneous regression, while others experiencing an aggressive, lethal disease. Cholesteatoma in children can be divided into two types: congenital and acquired. Identifying good prognostic markers is needed to help select patients who will require immediate surgical intervention. Matrix metalloproteinase-2 (MMP2) was previously reported to play an important role in cholesteatoma progression, by promoting bone destruction and keratinocyte infiltration. Herein, we analyzed MMP2 mRNA expression level in cholesteatoma using RNA-in situ hybridization in formalin-fixed, paraffin-embedded (FFPE) tissue samples. Sixty patients with cholesteatoma under 15 years old, who underwent their primary surgery at Aichi Medical University's Otolaryngology Department, were analyzed for MMP2 expression level, using RNA-in situ hybridization. There were no significant differences in MMP2 mRNA expression level between congenital cholesteatoma and acquired cholesteatomas. In congenital cholesteatoma, higher MMP2 signals were observed in the open type than in the closed type (p < 0.001). In acquired cholesteatoma, higher MMP2 signals were observed in the pars tensa than in the pars flaccida (p < 0.001). MMP2 mRNA expression level was almost exclusively found in the fibroblasts or in the inflammatory cells in the stroma, but not in the epithelium. Our study reveals that MMP2 mRNA expression level is strongly associated with the subtypes of cholesteatoma. The findings suggest that the level of expression of MMP2 mRNA may be related to the pathogenesis and aggressive features of cholesteatoma.

529: A phase 1b, randomized, double-blind, placebo-controlled, dose-escalation trial of CB-280, an arginase inhibitor, in patients with cystic fibrosis

Journal of Cystic Fibrosis

2021 Nov 01

Boas, S;Donaldson, S;McBennett, K;Liou, T;Howrylak, J;Johnson, L;Teneback, C;Dozor, A;Sawicki, G;Dumlao, J;Pan, A;Akella, L;Zhang, J;Carroll, S;Orford, K;Kuriakose, E;Mermis, J;
| DOI: 10.1016/S1569-1993(21)01953-6

Background: In CF, impaired nitric oxide (NO) production may contribute to impaired host antimicrobial defense, chronic airway infection, and compromised pulmonary function. L-arginine (Arg) is a required substrate of NO synthases for production of NO. Depletion of Arg by arginase, an abundant enzyme expressed and secreted into airways by neutrophils, contributes to NO deficiency. Clinical studies in CF patients have shown that administration of inhaled Arg improved fractional exhaled NO (FeNO) and trended toward improvement in FEV1. CB-280 is a potent, reversible, oral arginase inhibitor that generates sustained increases in systemic Arg. In preclinical studies in CF mouse models, CB-280 improved central airway resistance and decreased lung infection. We present the first 2 doseescalation cohorts of an ongoing phase 1b study of CB-280 versus placebo in adults with CF (NCT04279769; CX-280-202). Methods: Patients were randomized 3:1 to receive CB-280 or placebo orally over 14 days in 4 sequential dose-escalation cohorts (50, 100, 200, 400 mg twice a day; n = 8/cohort). Primary endpoint was safety and tolerability, as assessed by adverse events (AEs), laboratory changes, ECG, and spirometry. Results: Results are presented for 16 patients from the first 2 cohorts (n = 6 in 50-mg cohort, n = 6 in 100-mg cohort, n = 4 placebo). Median age was 31.5 and 69% were female. Median baseline ppFEV1 was 65%; 86% of patients were on elexacaftor/tezacaftor/ivacaftor at study entry. Treatmentemergent AEs occurred in 3 of 12 patients treated with CB-280 and 1 of 4 patients on placebo. Treatment-related AEs in CB-280-treated patients included Grade 1/2 dizziness and acne. No Grade 3 or greater events; doselimiting toxicities; serious AEs; or major adverse changes on lab assessments, ECG, or vital signs occurred in CB-280 patients. Spirometry showed no safety problems, and there was a positive trend in ppFEV1 with CB-280. CB-280 exhibited linear pharmacokinetics. Steady-state Ctrough at the 100-mg dose surpassed the IC90 for arginase inhibition in plasma, indicating continuous target coverage. Dose-related increases in plasma Arg were observed, with a mean 1.9-fold increase at the 100-mg dose. FeNO showed a slightly positive trend from baseline for patients on CB-280. Updated data will be presented. Conclusion: In conclusion, CB-280 was well tolerated in the initial dose cohorts of this study, with no CB-280 patients experiencing dose-limiting toxicities. CB-280 exhibited linear pharmacokinetics, achieving continuous target coverage in plasma at the 100-mg dose, and trended favorably in FEV1 and FeNO parameters.

Genetic Deletion of the Prostaglandin EP3 Receptor in the Kidney Tubule of Adult Mice Has No Impact on Kidney Water Handling

The FASEB Journal

2021 Jan 01

Esteva‐Font, C;Krogager, T;Hoorn, E;Fenton, R;
| DOI: 10.1096/fasebj.2021.35.S1.00332

Prostaglandin E2 (PGE2) is an important lipid mediator modulating various aspects of kidney function. PGE2 exerts its effects via four PGE2 receptors, EP1-EP4. The EP3 receptor is expressed in the thick ascending limb (TAL) and the collecting duct, where it is proposed to inhibit cAMP generation and NaCl and water reabsorption. However, EP3 is also expressed in endothelial cells of arteries and arterioles, which also play a role in kidney function. Therefore, to assess the tubular role of EP3 in adult mice we generated a mouse model based on the Pax8Cre system with doxycycline-dependent deletion of EP3 along the renal tubule and assessed their renal phenotype in respect to water handling. RNAscope confirmed that EP3 was highly expressed in cortical and medullary TAL and collecting ducts, but it was non-detected in proximal tubule and thin limbs. Two weeks after treatment with doxycycline, EP3 mRNA expression was reduced by >80% in whole kidney (RT-q-PCR) and non-detectable (RNAscope) in tubules of knockout mice compared to control mice. There were no compensatory changes in other EP receptors. Under basal conditions, there were no significant differences in food and water intake, bodyweight, urinary output or plasma and urine biochemistries in both male or female control and knockout mice. There were no differences between genotypes in their renal handling of water during an acute water load, or in their response to the vasopressin V2 receptor agonist dDAVP. Rats drinking 1% NaCl for several days in combination with an EP3 antagonist have increased urine output (Hao et al., 2016). However, relative to controls, we could not detect significant differences in urine volume or osmolality in tubular EP3 knockout mice during 1% NaCl intake. In conclusion, EP3 in the renal tubule is not important for renal water handling or compensatory mechanisms exist. This new model provides a novel tool for examination of the role of EP3 in other aspects of renal function or kidney disease independently of potential developmental abnormalities or systemic effects.

RF10 | PMON205 LH/CG Receptor Activation Protects Mice from Diet-Induced Obesity and Modifies Adipose Tissue Immune Response

Journal of the Endocrine Society

2022 Nov 01

Lizneva, D;Ievleva, K;Gumerova, A;Shelly, E;Korkmaz, F;Muradova, V;Netto, J;Kuo, T;Sultana, F;Kumar, P;Kramskiy, N;Ryu, V;Padilla, A;Hutchison, S;Yuen, T;Zaidi, M;
| DOI: 10.1210/jendso/bvac150.058

Menopause is associated with the loss of LH ovulatory surges and enhanced visceral adiposity. Visceral fat depots increase from 5-8% at premenopause to 15-20% of total body fat at postmenopause. Here, we report that high-dose LH, hCG, or small molecule LH/CGR agonist ORG43553 injected twice-a-week into 14-weeks-old C57BL/6 male mice protects them from diet-induced obesity. Testosterone levels were elevated in mice treated with LH or hCG, but not with ORG43553. Notably, the anti-obesity action of LH/hCG is independent of testosterone, as blocking the androgen receptor using flutamide yielded similar results. Importantly, male Lhcgr knockout mice on a high-fat diet treated with LH failed to display a reduction in adiposity, confirming the in vivo specificity of action. Furthermore, our data phenocopied Lhcgr haploinsufficiency in mice. We confirmed the presence of Lhcgr in mouse genital and inguinal fat pads, adipose-derived stromal vascular cells, as well as in differentiated and undifferentiated 3T3-L1 murine adipocytes by qPCR, RNAscope in situ hybridization, and immunohistochemistry. Sanger sequencing showed that the extracellular domain of Lhcgr in genital fat depot was identical to the ovarian receptor. Similarly, we identified LHCGR in human subcutaneous and visceral fat depots. Binding of intraperitoneally injected AlexaFluor-488-labeled hCG was found not only in mouse ovary, but also in genital and subcutaneous fat pad, further confirming the presence of LHCGR in adipose tissue. This binding could be competitively displaced in 3T3-L1 cells using unlabeled hCG. LH, hCG and ORG43553 activated ERK1/2 in a dose-dependent manner in undifferentiated and differentiated 3T3-L1 cells, suggesting that the adipose LHCGR is fully functional. LH, hCG, and ORG43553 reduced adipogenic differentiation in 3T3-L1 cells, which is further confirmed by RNA sequencing. Moreover, we observed, that LH and hCG also alters several aspects of immune response in adipose tissue, including inflammatory response and adaptive immunity. In conclusion, we demonstrated that LH/CG receptors are present and fully functional in adipose tissue, and that high-dose intermittent activation of LHCGR in mouse fat depots protects mice from diet-induced obesity and modifies adipose tissue immune response. Presentation: Saturday, June 11, 2022 1:42 p.m. - 1:47 p.m., Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.

The intestinal γδ T cells: functions in the gut and in the distant organs

Frontiers in Immunology

2023 Jun 16

Li, G;Xia, J;Zeng, W;Luo, W;Liu, L;Zeng, X;Cao, D;
| DOI: 10.3389/fimmu.2023.1206299

Located in the frontline against the largest population of microbiota, the intestinal mucosa of mammals has evolved to become an effective immune system. γδ T cells, a unique T cell subpopulation, are rare in circulation blood and lymphoid tissues, but rich in the intestinal mucosa, particularly in the epithelium. Via rapid production of cytokines and growth factors, intestinal γδ T cells are key contributors to epithelial homeostasis and immune surveillance of infection. Intriguingly, recent studies have revealed that the intestinal γδ T cells may play novel exciting functions ranging from epithelial plasticity and remodeling in response to carbohydrate diets to the recovery of ischemic stroke. In this review article, we update regulatory molecules newly defined in lymphopoiesis of the intestinal γδ T cells and their novel functions locally in the intestinal mucosa, such as epithelial remodeling, and distantly in pathological setting, e.g., ischemic brain injury repair, psychosocial stress responses, and fracture repair. The challenges and potential revenues in intestinal γδ T cell studies are discussed.

Examining Neuronal Subtype-Specific Genes Downregulated in Schizophrenia (SCZ) Using RNAscope in the Dorso-Lateral Prefrontal Cortex (DLPFC)

Biological Psychiatry

2021 May 01

Kim, D;Jambhale, A;Kim, P;Kim, B;Lipska, B;Auluck, P;Marenco, S;
| DOI: 10.1016/j.biopsych.2021.02.630

Background Bulk tissue RNA-sequencing studies in human post-mortem brains have reported differentially expressed genes between patients with Schizophrenia (SCZ) and controls. Single-nucleus RNA-sequencing data indicate that genes downregulated in SCZ were particularly enriched in a sub-population of excitatory neurons (Ex21) indexed by SMYD1 gene. POSTN and NR4A2 are two genes highly specific to Ex21 and significantly downregulated in SCZ. We hypothesized that either the number of Ex21 cells or the expression of POSTN and NR4A2 within them would be reduced in SCZ vs. controls. Methods We conducted a RNAscope in situ hybridization study in 20 SCZ and 20 controls. Three sections of the DLPFC per individual were stained for SMYD1 and NR4A2, and another three for SMYD1 and POSTN, using the colorimetric version of RNAscope (240 sections). Each RNAscope slide was counterstained using hematoxylin/eosin. Additional sections were stained with NeuN to assess neuronal numbers. Results ANCOVA revealed that there were no significant differences between SCZ and controls in the proportion of Ex21 (SMYD1+) cells (p = 0.40 for POSTN dataset, 0.36 for NR4A2 dataset), or the expression of marker and target genes within SMYD1+ cells in the DLPFC (p = 0.86 for POSTN dataset, p = 0.15 for NR4A2 dataset). Conclusions These results contradict prior conclusions that both POSTN and NR4A2 are downregulated in SCZ, but the modest sample size and several methodological issues suggest caution in interpreting these results.

Somatosensory neurons express specific sets of lincRNAs, and lincRNA CLAP promotes itch sensation in mice

EMBO reports

2022 Dec 16

Wang, B;Jiang, B;Li, GW;Dong, F;Luo, Z;Cai, B;Wei, M;Huang, J;Wang, K;Feng, X;Tong, F;Wang, S;Wang, Q;Han, Q;Li, C;Zhang, X;Yang, L;Bao, L;
PMID: 36524339 | DOI: 10.15252/embr.202154313

Somatosensory neurons are highly heterogeneous with distinct types of neural cells responding to specific stimuli. However, the distribution and roles of cell-type-specific long intergenic noncoding RNAs (lincRNAs) in somatosensory neurons remain largely unexplored. Here, by utilizing droplet-based single-cell RNA-seq (scRNA-seq) and full-length Smart-seq2, we show that lincRNAs, but not coding mRNAs, are enriched in specific types of mouse somatosensory neurons. Profiling of lincRNAs from single neurons located in dorsal root ganglia (DRG) identifies 200 lincRNAs localized in specific types or subtypes of somatosensory neurons. Among them, the conserved cell-type-specific lincRNA CLAP associates with pruritus and is abundantly expressed in somatostatin (SST)-positive neurons. CLAP knockdown reduces histamine-induced Ca2+ influx in cultured SST-positive neurons and in vivo reduces histamine-induced scratching in mice. In vivo knockdown of CLAP also decreases the expression of neuron-type-specific and itch-related genes in somatosensory neurons, and this partially depends on the RNA binding protein MSI2. Our data reveal a cell-type-specific landscape of lincRNAs and a function for CLAP in somatosensory neurons in sensory transmission.

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	Description
sense Example: Hs-LAG3-sense	Standard probes for RNA detection are in antisense. Sense probe is reverse complent to the corresponding antisense probe.
Intron# Example: Mm-Htt-intron2	Probe targets the indicated intron in the target gene, commonly used for pre-mRNA detection
Pool/Pan Example: Hs-CD3-pool (Hs-CD3D, Hs-CD3E, Hs-CD3G)	A mixture of multiple probe sets targeting multiple genes or transcripts
No-XSp Example: Hs-PDGFB-No-XMm	Does not cross detect with the species (Sp)
XSp Example: Rn-Pde9a-XMm	designed to cross detect with the species (Sp)
O# Example: Mm-Islr-O1	Alternative design targeting different regions of the same transcript or isoforms
CDS Example: Hs-SLC31A-CDS	Probe targets the protein-coding sequence only
EnEm	Probe targets exons n and m
En-Em	Probe targets region from exon n to exon m
Retired Nomenclature
tvn Example: Hs-LEPR-tv1	Designed to target transcript variant n
ORF Example: Hs-ACVRL1-ORF	Probe targets open reading frame
UTR Example: Hs-HTT-UTR-C3	Probe targets the untranslated region (non-protein-coding region) only
5UTR Example: Hs-GNRHR-5UTR	Probe targets the 5' untranslated region only
3UTR Example: Rn-Npy1r-3UTR	Probe targets the 3' untranslated region only
Pan Example: Pool	A mixture of multiple probe sets targeting multiple genes or transcripts

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