Capturing the Biologic Onset of Inflammatory Bowel Diseases: Impact on Translational and Clinical Science

While much progress has been made in the last two decades in the treatment and the management of inflammatory bowel diseases (IBD)-both ulcerative colitis (UC) and Crohn's Disease (CD)-as of today these conditions are still diagnosed only after they have become symptomatic. This is a major drawback since by then the inflammatory process has often already caused considerable damage and the disease might have become partially or totally unresponsive to medical therapy. Late diagnosis in IBD is due to the lack of accurate, non-invasive indicators that would allow disease identification during the pre-clinical stage-as it is often done in many other medical conditions. Here, we will discuss what is known about the biologic onset and pre-clinical CD with an emphasis on studies conducted in patients' first degree relatives. We will then review the possible strategies to diagnose IBD very early in time including screening, available disease markers and imaging, and the possible clinical implications of treating these conditions at or close to their biologic onset. Later, we will review the potential impact of conducting translational research in IBD during the pre-clinical stage, especially focusing on the role of the microbiome in disease etiology and pathogenesis. Finally, we will highlight possible future developments in the field and how they can impact IBD management and our scientific knowledge of these conditions

Helicobacter Pylori-Negative MALT Lymphoma: A Series of Two Cases Presenting with Life-Threatening Upper Gastrointestinal Bleeding

Mucosa-associated lymphoid tissue (MALT) lymphoma is a common cause of gastric lymphoma. Although most cases are associated with an H. pylori infection, approximately 10% are H. pylori-negative. Patients with gastric MALT lymphoma are usually asymptomatic or present with nonspecific symptoms such as abdominal pain, dyspepsia, weight loss, and occult gastrointestinal bleeding. In this report, we describe two patients with H. pylori-negative MALT lymphoma who both presented with acute upper gastrointestinal bleeding that led to hemodynamic instability. After resuscitation, emergent endoscopy was performed. Both patients had the t (11; 18) (q21; q21) translocation, which prompted direct treatment by radiotherapy

High-Throughput Multi-Analyte Luminex Profiling Implicates Eotaxin-1 in Ulcerative Colitis

<div><p>Accurate and high-throughput technologies are needed for identification of new therapeutic targets and for optimizing therapy in inflammatory bowel disease. Our aim was to assess multi-analyte protein-based assays of cytokines/chemokines using Luminex technology. We have reported that Luminex-based profiling was useful in assessing response to L-arginine therapy in the mouse model of dextran sulfate sodium colitis. Therefore, we studied prospectively collected samples from ulcerative colitis (UC) patients and control subjects. Serum, colon biopsies, and clinical information were obtained from subjects undergoing colonoscopy for evaluation of UC or for non-UC indications. In total, 38 normal controls and 137 UC cases completed the study. Histologic disease severity and the Mayo Disease Activity Index (DAI) were assessed. Serum and colonic tissue cytokine/chemokine profiles were measured by Luminex-based multiplex testing of 42 analytes. Only eotaxin-1 and G-CSF were increased in serum of patients with histologically active UC vs. controls. While 13 cytokines/chemokines were increased in active UC vs. controls in tissues, only eotaxin-1 was increased in all levels of active disease in both serum and tissue. In tissues, eotaxin-1 correlated with the DAI and with eosinophil counts. Increased eotaxin-1 levels were confirmed by real-time PCR. Tissue eotaxin-1 levels were also increased in experimental murine colitis induced by dextran sulfate sodium, oxazolone, or <i>Citrobacter rodentium</i>, but not in murine <i>Helicobacter pylori</i> infection. Our data implicate eotaxin-1 as an etiologic factor and therapeutic target in UC, and indicate that Luminex-based assays may be useful to assess IBD pathogenesis and to select patients for anti-cytokine/chemokine therapies.</p></div

Tissue cytokines/chemokines profiles.

<p>± SD. Statistical analysis performed as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0082300#pone-0082300-t002" target="_blank">Table 2</a>. *<i>p</i><0.05, **<i>p</i><0.01, ***<i>p</i><0.001 vs. control; §<i>p</i><0.05, §§<i>p</i><0.01, §§§<i>p</i><0.001 vs. quiescent UC. ^a<i>n</i> = 34 controls, 26 quiescent UC, and 60 active UC. ^b<i>n</i> = 20 controls, 19 quiescent UC, and 23 active UC. ^c<i>n</i> = 34 controls, 26 quiescent UC, and 60 active UC. UC activity determined by histology. Values are in pg/mg protein. Mean </p

Colon tissue eotaxin-1 is significantly increased in UC.

<p>Snap frozen colonic biopsies were lysed and cytokine/chemokine levels were measured by Luminex technology, with each sample corrected for tissue lysate protein concentration, all as described in the Methods. (A) Eotaxin-1 and (B) G-CSF concentration in control and all UC patients. (C) Eotaxin-1 and (D) G-CSF concentration in UC tissues stratified by histologic disease severity, and in control subjects. <i>n</i> = 34 for control, and <i>n</i> = 86 for UC. **<i>p</i><0.01; ***<i>p</i><0.001 vs. control. ^#<i>p</i><0.05; ^##<i>p</i><0.01 vs. quiescent colitis. §<i>p</i><0.05 vs. mild colitis.</p

Eotaxin-1 mRNA expression is increased in UC tissues vs. control subjects and in involved vs. uninvolved tissues in UC patients with colitis limited to the left colon.

<p>Tissue samples were snap frozen, and subsequently RNA was extracted and mRNA expression assessed by real-time PCR. (A) Eotaxin-1 mRNA levels in control and all UC patients. (B) Eotaxin-1 mRNA levels stratified by UC histologic disease severity. <i>n</i> = 22 for controls, and <i>n</i> = 46 for UC patients. **<i>p</i><0.01; ***<i>p</i><0.001 vs. control. ^###<i>p</i><0.001 vs. quiescent colitis. §<i>p</i><0.05 vs. mild colitis. (C) Eotaxin-1 mRNA expression in paired uninvolved (right-sided) tissue and involved (left-sided) tissues. <i>n</i> = 21 paired samples from UC patients. ***<i>p</i><0.001 vs. paired uninvolved tissue.</p

Colonic tissue eotaxin-1 levels are increased in mouse models of colitis but not in <i>H. pylori</i>-induced gastritis.

<p>Tissue samples were obtained at the time of sacrifice, placed in RIPA buffer, then lysed. Eotaxin-1 levels were assessed by Luminex assay. Each sample was corrected for tissue lysate protein concentration. Tissues were obtained as follows: (A) After 7 days of 4% DSS in the drinking water. (B) 3 days after intrarectal exposure to oxazolone. (C) 14 days after initiation of <i>C. rodentium</i> infection. (D) 4 months after inoculation with <i>H. pylori</i>. *<i>p</i><0.05; **<i>p</i><0.01 vs. control.</p

Eotaxin-2 and -3 levels in UC.

<p>Serum and tissue cytokine/chemokine levels are based on Luminex assay using a custom kit for eotaxin-2 and eotaxin-3. (A – B) Serum levels. (C – D) Tissue levels. For serum, <i>n</i> = 33 for control and <i>n</i> = 101 for UC. For tissue, <i>n</i> = 18 for control and <i>n</i> = 50 for UC. Note that many samples did not have reliable eotaxin-3 results.</p