An Enriched Environment Reduces Chronic Stress-Induced Visceral Pain Through Modulating Microglial Activity in the Central Nucleus of the Amygdala

American journal of physiology. Gastrointestinal and liver physiology

2021 Dec 08

Yuan, T;Orock, A;Greenwood-Van Meerveld, B;
PMID: 34877892 | DOI: 10.1152/ajpgi.00307.2021

Cognitive behavioral therapy (CBT) improves the quality of life for patients with brain-gut disorders, however, the underlying mechanisms of CBT remain to be explored. Previously we showed that environmental enrichment (EE), an experimental paradigm that mirrors positive behavioral intervention, ameliorates chronic stress-induced visceral hypersensitivity in a rodent model via mechanisms involving altered activity in the central nucleus of amygdala (CeA). In the present study, we investigated whether microglia-mediated synaptic plasticity in the CeA is a potential mechanism underlying the protective effects of EE against stress-induced visceral hypersensitivity. We sterotaxically implanted corticosterone (CORT) micropellets onto the dorsal margin of the CeA shown previously to induce colonic hypersensitivity. Animals were housed in EE cages or standard cages for 14 days following CORT implantation. Visceral sensitivity was assessed via visceromotor behavioral response to colorectal distension. Microglial morphology, microglia-mediated synaptic engulfment and the expression of synaptic pruning-related signals C1q, C3 and C3R were measured using immunofluorescence and RNAscope assay. We found that housing CORT implanted rats in EE cages for 14 days attenuated visceral hypersensitivity in both male and female rats as compared to control rats maintained in standard housing. EE reduced CORT-induced microglial remodeling and microglia-mediated synaptic pruning with reduced C1q and CR3, but not C3, expression. Our data suggest that exposure to EE is sufficient to ameliorate stress-induced visceral pain via reducing amygdala microglia-modulated neuronal plasticity.

Circulating Monocytes Associated with Anti-PD-1 Resistance in Human Biliary Cancer Induce T Cell Paralysis

SSRN Electronic Journal

2021 Dec 21

Keenan, B;McCarthy, E;Ilano, A;Yang, H;Zhang, L;Allaire, K;Fan, Z;Li, T;Lee, D;Sun, Y;Cheung, A;Luong, D;Chang, H;Marquez, J;Sheldon, B;Kelley, R;ye, j;Fong, L;
| DOI: 10.2139/ssrn.3985187

Although suppressive myeloid cells have been proposed as a mechanism of resistance to immunotherapy, their role in response to checkpoint inhibitor treatment (CPI) in anti-PD-1 refractory cancers, such as biliary tract cancer (BTC), is largely unknown. We used multiplexed single-cell transcriptomic and epitope sequencing to profile >200,000 peripheral blood mononuclear cells from advanced BTC. In BTC patients, CD14+ monocytes expressing high levels of immunosuppressive cytokines and trafficking molecules involved in chemotaxis (CD14CTX) are associated with resistance to CPI. CD14CTX can directly suppress CD4+ T cells and induce SOCS3 expression in naive CD4+ T cells rendering them functionally unresponsive. Gene signatures from CD14CTX are correlated with worse survival in BTC patients as well as in other anti-PD-1 refractory cancers. These results demonstrate that monocytes arising in the setting of anti-PD-1 insensitivity can induce T cell paralysis as a distinct mode of tumor-mediated immunosuppression.

Innovations in Placental Pathology

Benirschke's Pathology of the Human Placenta

2021 Dec 08

Pantham, P;Soncin, F;Zhang-Rutledge, K;Srinivasan, S;Lamale-Smith, L;Laurent, L;Parast, M;
| DOI: 10.1007/978-3-030-84725-8_31

Over the past two decades, multiple new methods have been developed for probing the structure and function of human tissues and organ systems. These innovative methods have paved the road toward a new era in medicine, where diseases are subclassified, not only based on histology but also at the molecular level, and often based on an integrated assessment of clinical signs and symptoms, detailed in vivo imaging, and histologic and molecular evaluation of biopsied tissues. Advances in detection, identification, and quantification of both cell-free and extracellular vesicle-derived biomarkers in the blood are more commonly allowing for noninvasive measures of organ function, precluding the need for invasive biopsies. In addition, stem cells, “organoids,” and “organ-on-a-chip” models have permitted the study of human cells and tissues in greater detail and complexity. The application of these innovative methods to the human placenta is detailed in this chapter, highlighting not only the new information gained about the structure and function of this important transient organ but also the ways in which this information can be translated into, and thus transform, the practice of placental pathology.

Ulcerative colitis: shedding light on emerging agents and strategies in preclinical and early clinical development

Expert opinion on investigational drugs

2021 Aug 10

Caballol, B;Gudiño, V;Panes, J;Salas, A;
PMID: 34365869 | DOI: 10.1080/13543784.2021.1965122

Ulcerative colitis (UC) is an inflammatory disease of the large intestine. Progress in preclinical therapeutic target discovery and clinical trial design has resulted in the approval of new therapies. Nonetheless, remission rates remain below 30% thus underlining the need for novel, more effective therapies.This paper reviews current experimental techniques available for drug testing in intestinal inflammation and examines new therapies in clinical development for the treatment of UC. The authors searched the literature for 'ulcerative colitis' AND 'preclinical' OR 'drug target/drug name' (i.e. infliximab, vedolizumab, IL-12, IL-23, JAK, etc.). Studies that included preclinical in vivo or in vitro experiments are discussed. The clinicaltrial.gov site was searched for 'ulcerative colitis' AND 'Recruiting' OR 'Active, not recruiting' AND 'Interventional (Clinical Trial)' AND 'early phase 1' OR 'phase 1' OR 'phase 2' OR 'phase 3.'Using in vivo, ex vivo, and/or in vitro models could increase the success rates of drugs moving to clinical trials, and hence increase the efficiency of this costly process. Selective JAK1 inhibitors, S1P modulators, and anti-p19 antibodies are the most promising options to improve treatment effectiveness. The development of drugs with gut-restricted exposure may provide increased efficacy and an improved safety.

Highly multiplexed mass cytometry identifies the immunophenotype in the skin of Dermatomyositis

The Journal of investigative dermatology

2021 Mar 22

Patel, J;Maddukuri, S;Li, Y;Bax, C;Werth, VP;
PMID: 33766508 | DOI: 10.1016/j.jid.2021.02.748

Dermatomyositis (DM) is a rare, systemic autoimmune disease that most frequently affects the skin, muscles, and lungs. The inflammatory infiltrate in skin has not been fully characterized and in this study we took an single cell, unbiased approach by using Imaging mass cytometry (IMC). Substantial monocyte-macrophage diversity was observed with the CD14+ population correlated positively with cutaneous dermatomyositis disease area and severity index (CDASI) scores (p=0.031). The T cell compartment revealed CD4+ T, CD8+ T, and FOXP3+ T cells. Activated (CD69+) circulating memory T cells correlated positively with CDASI scores (p=0.0268). IFNβ protein was highly upregulated in the T cell, macrophage, dendritic cell, and endothelial cell populations of DM skin. Myeloid DCs (mDCs) expressed pPPARγ, pIRF3, IL4, and IL31 and their quantity correlated with itch as measured in the Skindex-29. Plasmacytoid DCs (pDCs) colocalized with IFNγ in addition to the known colocalization with IFNβ. Nuclear pPPARγ expression was found in the DM endothelium. IMC allows us to characterize single cells in the immune cell population and identify upregulated cytokines and inflammatory pathways in DM. These findings have important implications for the development of future targeted therapies for DM.

Annexin A1 alleviates kidney injury by promoting the resolution of inflammation in diabetic nephropathy

Kidney international

2021 Mar 03

Wu, L;Liu, C;Chang, DY;Zhan, R;Sun, J;Cui, SH;Eddy, S;Nair, V;Tanner, E;Brosius, FC;Looker, HC;Nelson, RG;Kretzler, M;Wang, JC;Xu, M;Ju, W;Zhao, MH;Chen, M;Zheng, L;
PMID: 33675846 | DOI: 10.1016/j.kint.2021.02.025

Since failed resolution of inflammation is a major contributor to the progression of diabetic nephropathy, identifying endogenously generated molecules that promote the physiological resolution of inflammation may be a promising therapeutic approach for this disease. Annexin A1 (ANXA1), as an endogenous mediator, plays an important role in resolving inflammation. Whether ANXA1 could affect established diabetic nephropathy through modulating inflammatory states remains largely unknown. In the current study, we found that in patients with diabetic nephropathy, the levels of ANXA1 were upregulated in kidneys, and correlated with kidney function as well as kidney outcomes. Therefore, the role of endogenous ANXA1 in mouse models of diabetic nephropathy was further evaluated. ANXA1 deficiency exacerbated kidney injuries, exhibiting more severe albuminuria, mesangial matrix expansion, tubulointerstitial lesions, kidney inflammation and fibrosis in high fat diet/streptozotocin-induced-diabetic mice. Consistently, ANXA1 overexpression ameliorated kidney injuries in mice with diabetic nephropathy. Additionally, we found Ac2-26 (an ANXA1 mimetic peptide) had therapeutic potential for alleviating kidney injuries in db/db mice and diabetic Anxa1 knockout mice. Mechanistic studies demonstrated that intracellular ANXA1 bound to the transcription factor NF-κB p65 subunit, inhibiting its activation thereby modulating the inflammatory state. Thus, our data indicate that ANXA1 may be a promising therapeutic approach to treating and reversing diabetic nephropathy.

Using a Reporter Mouse to Map Known and Novel Sites of GLP-1 Receptor Expression in Peripheral Tissues of Male Mice

Endocrinology

2021 Mar 01

Andersen, DB;Grunddal, KV;Pedersen, J;Kuhre, RE;Lund, ML;Holst, JJ;Ørskov, C;
PMID: 33508122 | DOI: 10.1210/endocr/bqaa246

Glucagon-like peptide-1 receptor (GLP-1R) activation is used in the treatment of diabetes and obesity; however, GLP-1 induces many other physiological effects with unclear mechanisms of action. To identify the cellular targets of GLP-1 and GLP-1 analogues, we generated a Glp1r.tdTomato reporter mouse expressing the reporter protein, tdTomato, in Glp1r-expressing cells. The reporter signal is expressed in all cells where GLP-1R promoter was ever active. To complement this, we histologically mapped tdTomato-fluorescence, and performed Glp-1r mRNA in situ hybridization and GLP-1R immunohistochemistry on the same tissues. In male mice, we found tdTomato signal in mucus neck, chief, and parietal cells of the stomach; Brunner's glands; small intestinal enteroendocrine cells and intraepithelial lymphocytes; and myenteric plexus nerve fibers throughout the gastrointestinal tract. Pancreatic acinar-, β-, and δ cells, but rarely α cells, were tdTomato-positive, as were renal arteriolar smooth muscle cells; endothelial cells of the liver, portal vein, and endocardium; aortal tunica media; and lung type 1 and type 2 pneumocytes. Some thyroid follicular and parafollicular cells displayed tdTomato expression, as did tracheal cartilage chondrocytes, skin fibroblasts, and sublingual gland mucus cells. In conclusion, our reporter mouse is a powerful tool for mapping known and novel sites of GLP-1R expression in the mouse, thus enhancing our understanding of the many target cells and effects of GLP-1 and GLP-1R agonists.

Inverse correlation between PD-L1 expression and LGR5 expression in tumor budding of stage II/III colorectal cancer

Annals of Diagnostic Pathology

2021 Mar 01

Sato, K;Uehara, T;Nakajima, T;Iwaya, M;Miyagawa, Y;Watanabe, T;Ota, H;
| DOI: 10.1016/j.anndiagpath.2021.151739

We investigated the expression of LGR5, the most robust and reliable known cancer stem cell (CSC) marker of colorectal cancer, and PD-L1 in tumor budding (TB), as well as clinicopathological features. Tissue microarrays (TMAs) were generated from TB samples from 32 stage II/III colorectal adenocarcinoma patients, and LGR5 expression in TMAs was evaluated by RNAscope, an extremely sensitive RNA in situ hybridization technique. LGR5 expression was significantly lower in the PD-L1-positive group than in the PD-L1-negative group (P = 0.0256). In the PD-L1-positive group, the tumor-infiltrating lymphocytes (TILs) score tended to be higher while the TNM stage was lower compared with the PD-L1 negative group (P = 0.0822 and P = 0.0765, respectively). There was no significant difference in Overall Survival between the PD-L1-positive and PD-L1-negative groups (log-rank test, P = 0.8218). This study showed that PD-L1-positive patients are a unique population with low LGR5 expression, and that LGR5-positive cells may be a promising therapeutic target in PD-L1-negative patients.

Identification of Potential Meniere\'s Disease Targets in the Adult Stria Vascularis

Frontiers in neurology

2021 Feb 05

Gu, S;Olszewski, R;Nelson, L;Gallego-Martinez, A;Lopez-Escamez, JA;Hoa, M;
PMID: 33613436 | DOI: 10.3389/fneur.2021.630561

The stria vascularis generates the endocochlear potential and is involved in processes that underlie ionic homeostasis in the cochlear endolymph, both which play essential roles in hearing. The histological hallmark of Meniere's disease (MD) is endolymphatic hydrops, which refers to the bulging or expansion of the scala media, which is the endolymph-containing compartment of the cochlea. This histologic hallmark suggests that processes that disrupt ion homeostasis or potentially endocochlear potential may underlie MD. While treatments exist for vestibular symptoms related to MD, effective therapies for hearing fluctuation and hearing loss seen in MD remain elusive. Understanding the potential cell types involved in MD may inform the creation of disease mouse models and provide insight into underlying mechanisms and potential therapeutic targets. For these reasons, we compare published datasets related to MD in humans with our previously published adult mouse stria vascularis single-cell and single-nucleus RNA-Seq datasets to implicate potentially involved stria vascularis (SV) cell types in MD. Finally, we provide support for these implicated cell types by demonstrating co-expression of select candidate genes for MD within SV cell types.

Novel variants in the stem cell niche factor WNT2B define the disease phenotype as a congenital enteropathy with ocular dysgenesis

European journal of human genetics : EJHG

2021 Feb 01

Zhang, YJ;Jimenez, L;Azova, S;Kremen, J;Chan, YM;Elhusseiny, AM;Saeed, H;Goldsmith, J;Al-Ibraheemi, A;O'Connell, AE;Kovbasnjuk, O;Rodan, L;Agrawal, PB;Thiagarajah, JR;
PMID: 33526876 | DOI: 10.1038/s41431-021-00812-1

WNT2B is a member of the Wnt family, a group of signal transduction proteins involved in embryologic development and stem cell renewal and maintenance. We recently reported homozygous nonsense variants in WNT2B in three individuals with severe, neonatal-onset diarrhea, and intestinal failure. Here we present a fourth case, from a separate family, with neonatal diarrhea associated with novel compound heterozygous WNT2B variants. One of the two variants was a frameshift variant (c.423del [p.Phe141fs]), while the other was a missense change (c.722 G > A [p.G241D]) that we predict through homology modeling to be deleterious, disrupting post-translational acylation. This patient presented as a neonate with severe diet-induced (osmotic) diarrhea and growth failure resulting in dependence on parenteral nutrition. Her gastrointestinal histology revealed abnormal cellular architecture particularly in the stomach and colon, including oxyntic atrophy, abnormal distribution of enteroendocrine cells, and a paucity of colonic crypt glands. In addition to her gastrointestinal findings, she had bilateral corneal clouding and atypical genital development later identified as a testicular 46,XX difference/disorder of sexual development. Upon review of the previously reported cases, two others also had anterior segment ocular anomalies though none had atypical genital development. This growing case series suggests that variants in WNT2B are associated with an oculo-intestinal (and possibly gonadal) syndrome, due to the protein's putative involvement in multiple developmental and stem cell maintenance pathways.

Serotonin receptor 4 in the hippocampus modulates mood and anxiety

Molecular psychiatry

2021 Jan 13

Karayol, R;Medrihan, L;Warner-Schmidt, JL;Fait, BW;Rao, MN;Holzner, EB;Greengard, P;Heintz, N;Schmidt, EF;
PMID: 33441982 | DOI: 10.1038/s41380-020-00994-y

Serotonin receptor 4 (5-HT4R) plays an important role in regulating mood, anxiety, and cognition, and drugs that activate this receptor have fast-acting antidepressant (AD)-like effects in preclinical models. However, 5-HT4R is widely expressed throughout the central nervous system (CNS) and periphery, making it difficult to pinpoint the cell types and circuits underlying its effects. Therefore, we generated a Cre-dependent 5-HT4R knockout mouse line to dissect the function of 5-HT4R in specific brain regions and cell types. We show that the loss of functional 5-HT4R specifically from excitatory neurons of hippocampus led to robust AD-like behavioral responses and an elevation in baseline anxiety. 5-HT4R was necessary to maintain the proper excitability of dentate gyrus (DG) granule cells and cell type-specific molecular profiling revealed a dysregulation of genes necessary for normal neural function and plasticity in cells lacking 5-HT4R. These adaptations were accompanied by an increase in the number of immature neurons in ventral, but not dorsal, dentate gyrus, indicating a broad impact of 5-HT4R loss on the local cellular environment. This study is the first to use conditional genetic targeting to demonstrate a direct role for hippocampal 5-HT4R signaling in modulating mood and anxiety. Our findings also underscore the need for cell type-based approaches to elucidate the complex action of neuromodulatory systems on distinct neural circuits.

LncRNA DANCR represses Doxorubicin-induced apoptosis through stabilizing MALAT1 expression in colorectal cancer cells

Cell death & disease

2021 Jan 06

Xiong, M;Wu, M;Dan Peng, ;Huang, W;Chen, Z;Ke, H;Chen, Z;Song, W;Zhao, Y;Xiang, AP;Zhong, X;
PMID: 33414433 | DOI: 10.1038/s41419-020-03318-8

Long non-coding RNA (lncRNA) DANCR has been reported to participate in key processes such as stem cell differentiation and tumorigenesis. In a high throughput screening for lncRNAs involved in Doxorubicin-induced apoptosis, we found DANCR was suppressed by Doxorubicin and it acted as an important repressor of apoptosis in colorectal cancer. Further studies demonstrated that DANCR promoted the oncogenic lncRNA MALAT1 expression via enhancing the RNA stability of MALAT1 to suppress apoptosis. MALAT1 could efficiently mediate the suppressive function of DANCR on apoptosis. Mechanistic studies found the RNA-binding protein QK served as an interacting partner of both DANCR and MALAT1, and the protein level of QK was subjected to the regulation by DANCR. Furthermore, QK was able to modulate the RNA stability of MALAT1, and the interaction between QK and MALAT1 was controlled by DANCR. In addition, QK could mediate the function of DANCR in regulating the expression of MALAT1 and suppressing apoptosis. These results revealed DANCR played a critical role in Doxorubicin-induced apoptosis in colorectal cancer cells, which was achieved by the interaction between DANCR and QK to enhance the expression of MALAT1.

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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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