CD44 Upregulation in E-Cadherin-Negative Esophageal Cancers Results in Cell Invasion

E-cadherin is frequently lost during epithelial-mesenchymal transition and the progression of epithelial tumorigenesis. We found a marker of epithelial-mesenchymal transition, CD44, upregulated in response to functional loss of E-cadherin in esophageal cell lines and cancer. Loss of E-cadherin expression correlates with increased expression of CD44 standard isoform. Using an organotypic reconstruct model, we show increased CD44 expression in areas of cell invasion is associated with MMP-9 at the leading edge. Moreover, Activin A increases cell invasion through CD44 upregulation after E-cadherin loss. Taken together, our results provide functional evidence of CD44 upregulation in esophageal cancer invasion

Association Of Tgfβ Signaling With The Maintenance Of A Quiescent Stem Cell Niche In Human Oral Mucosa

A dogma in squamous epithelial biology is that proliferation occurs in the basal cell layer. Notable exceptions are squamous epithelia of the human oral cavity, esophagus, ectocervix, and vagina. In these human epithelia, proliferation is rare in the basal cell layer, and the vast majority of cells positive for Ki67 and other proliferation markers are found in para- and suprabasal cell layers. This unique human feature of a generally quiescent basal cell layer overlaid by highly proliferative cells offers the rare opportunity to study the molecular features of undifferentiated, quiescent, putative stem cells in their natural context. Here, we show that the quiescent human oral mucosa basal cell layer expresses putative markers of stemness, while para- and suprabasal cells are characterized by cell cycle genes. We identified a TGFβ signature in this quiescent basal cell layer. In in vitro organotypic cultures, human keratinocytes could be induced to express markers of these quiescent basal cells when TGFβ signaling is activated. The study suggests that the separation of basal cell layer and proliferation in human oral mucosa may function to accommodate high proliferation rates and the protection of a quiescent reserve stem cell pool. Psoriasis, an epidermal inflammatory hyperproliferative disease, exhibits features of a quiescent basal cell layer mimicking normal oral mucosa. Our data indicate that structural changes in the organization of epithelial proliferation could contribute to longevity and carcinogenesis

Inverse correlation of E-cadherin and CD44 expression in primary human esophageal tumors.

<p>IHC with anti-CD44 antibody shows increased CD44 expression during the progression from normal (A) to carcinoma <i>in situ,</i> CIS (B), and tumor (C). Antibody against E-cadherin and CD44 show expression of E-cadherin (D) and in the absence of CD44 (E) in the normal epithelium. In EAC tissues with retained E-cadherin expression (F, dashed black line) the signal for CD44 is low (G). (H) Loss of E-cadherin is associated with an intense signal for CD44 (I). Scale bar is 50 micron.</p

CD44 co-localizes with MMP-9 in areas of invasion and is upregulated by Activin A.

<p>(A) For <i>in situ</i> zymography, areas of MMP activity are highlighted by a positive fluorescein signal on frozen sections of Ecad, EC and ECdnT organotypic cultures. (B) Zymography using organotypic culture conditioned medium collected from Ecad, EC and ECdnT cells illustrates increased secretion of MMP-2 and MMP-9. (C) Double immunofluorescence staining of Ecad, EC and ECdnT shows MMP-9 (red) colocalization with CD44 (green) in invasive cells. Scale bar is 50 micron. (D) Stimulation with Activin A induces enhanced invasion of ECdnT and TE-11 cells in invasion assays. Comparison between groups was done by Man and Whitney non- parametrical test, * p<0.05, **p<0.001. (E) Western Blot of cell lysates from TE-11 transfected with AllStars negative non-silencing siRNA cells (ctrl), and TE-11 cells transfected with siRNA against E-cadherin (si1, si2) demonstrates upregulation of CD44 after stimulation with Activin A.</p

CD44 expression is increased in areas of invasion after E-cadherin loss.

<p>Immunohistochemistry of paraffin sections with anti-E-cadherin antibody (A), anti-TGFβRII (B) and anti-CD44 (C) shows CD44–positive cells in EC and ECdnT cells invading into the underlying collagen/matrigel matrix (arrows) that are negative for E-cadherin and TGFβRII. Immunohistochemical staining for the EMT markers S100A4 (D), vimentin (E) and αSMA (F) shows increased positive signal in invading cells. Scale bar is 50 micron.</p

Expression of CD44, TGFβRII, vimentin and E-cadherin in cancer cell lines.

<p>(A) Western Blot analysis of esophageal squamous carcinoma cell lines (TE-1 -7, -8, -11, and -12), esophageal adenocarcinoma cell lines (FLO-1, OE33, SK-GT-4) and head and neck cancer cell lines (JHU-012, JHU-013) show that expression of E-cadherin and TGFβRII is associated with low levels of CD44, while loss of E-cadherin and TGFβRII correlates with up-regulation of CD44 and the EMT marker vimentin. (B) Restoration of E-cadherin, Ecad, expression by retroviral transfection of TE-8 and FLO-1 cells compared to vector control (neo) demonstrates an overall decrease in CD44 expression along with a switch to increased CD44v over CD44s (CD44v: CD44variant, CD44s: CD44 standard).</p

Influence of prior intravenous thrombolysis in patients treated with mechanical thrombectomy for M2 occlusions: insight from the Endovascular Treatment in Ischemic Stroke (ETIS) registry.

International audienceBackground Intravenous thrombolysis (IVT) for patients treated with mechanical thrombectomy (MT) for proximal occlusions has recently been questioned through randomized trials. However, few patients with M2 occlusions were included. We investigated the influence of prior IVT for patients presenting M2 occlusions treated with MT in comparison with MT alone.Methods We conducted a retrospective analysis of the Endovascular Treatment in Ischemic Stroke (ETIS) registry, a multicenter observational study. Data from consecutive patients treated with MT for M2 occlusions between January 2015 and January 2022 at 26 comprehensive stroke centers were analyzed. The primary endpoint was 90-day modified Rankin Scale score of 0–2. Outcomes were compared using propensity score approaches. We also performed sensitivity analysis in relevant subgroups of patients.Results Among 1132 patients with M2 occlusions treated with MT, 570 received prior IVT. The two groups were comparable after propensity analysis. The rate of favorable functional outcome was significantly higher in the IVT+MT group compared with the MT alone group (59.8% vs 44.7%; adjusted OR 1.38, 95% CI 1.10 to 1.75, P=0.008). Hemorrhagic and procedural complications were similar in both groups. In sensitivity analysis excluding patients with anticoagulation treatment, favorable recanalization was more frequent in the IVT+MT group (OR 1.37, 95% CI 1.11 to 1.70, P=0.004).Conclusions In cases of M2 occlusions, prior IVT combined with MT resulted in better functional outcome than MT alone, without increasing the rate of hemorrhagic or procedural complications. These results suggest the benefit of IVT in patients undergoing MT for M2 occlusions

Mechanical thrombectomy practices in France: Exhaustive survey of centers and individual operators

International audienc