LRIG1 as a Potential Novel Marker for Neoplastic Transformation in Ocular Surface Squamous Neoplasia

<div><p>The leucine rich repeats and immunoglobulin-like protein 1 (LRIG1) is a newly discovered negative regulator of epidermal growth factor receptor (EGFR) and a proposed tumor suppressor. It is not universally downregulated in human cancers, and its role in neoplastic transformation and tumorigenesis is not well-documented. In this study, we show the expression of LRIG1 as a novel potential marker for neoplastic transformation in ocular-surface squamous neoplasia (OSSN). The following two groups were included in this study: 1) benign group (3 cases; 1 with papilloma and 2 with dysplasia) and 2) malignant group (3 cases with squamous cell carcinoma (SCC)). In both groups, immunofluorescence analysis was firstly performed for keratins 4, 12, 13, and 15 to characterize the state of differentiation, and for Ki67 to evaluate the proliferation activity. Subsequently, LRIG1 and EGFR expression was analyzed. Either keratin 4 and/or 13, both non-keratinized epithelial cell markers, were generally expressed in both groups, except for 1 severe SCC case. Keratin 15, an undifferentiated basal cell marker, was more strongly expressed in the malignant cases than in the benign cases. The Ki67 index was significantly higher (<i>P</i><0.002) in the malignant group (33.2%) than in the benign group (10.9%). LRIG1 expression was limited to basal epithelial cells in normal corneal epithelial tissue. Interestingly, LRIG1 was expressed throughout the epithelium in all the benign cases. In contrast, its expression was limited or totally disappeared in the malignant cases. Inversely, EGFR staining was faintly expressed in the benign cases, yet strongly expressed in the malignant cases. Malignant tissue with proliferative potential presented EGFR overexpression and inverse downregulation of LRIG1, consistent with LRIG1 being a suppressor of neoplastic transformation by counteracting the tumor growth property of EGFR. Our findings indicate that downregulation of LRIG1 is possibly a novel potential marker of transformation and tumorigenesis in OSSN cases.</p></div

Clinical presentation of 6 cases with OSSN.

<p>(A) Case 1 with papilloma. Multiple sessile masses were prominent in lower conjunctival fornix and palpebral conjunctiva (arrows). (B) Case 2 with severe dysplasia. Elevated smooth mass lesion in nasal side of limbus to bulbar conjunctiva (arrows) presented gelatinous surface with typical corkscrew vascular tufts. (C) Case 3 with severe dysplasia. Irregularly elevated mass was diffusely covered whole ocular surface. Flat gelatinous lesion and irregular whitish lesion (arrows) were mixed. Lightly pigmented area was also observed (arrowheads). (D) Case 4 with CIS. Elevated nodular mass with vascular tufting was covered temporal side of limbus to bulbar conjunctiva. (E) Case 5 with CIS. Nodular lesion with typical vascular tufts extended from the caruncle to bulbar conjunctiva. (F) Case 6 with invasive SCC. Gelatinous irregular epithelium covered whole ocular surface.</p

The expression pattern of LRIG1 and EGFR.

<p>The expression pattern of LRIG1 (A–C, G–I) and EGFR (D–F, J–L) in case 1 (A, D), case 2 (B, E), case 3 (C, F), case 4 (G, J), case 5 (H, K), and case 6 (I, L). The expression pattern in conjunctivas (a, d), limbus (b, e), and corneas (c, f) in healthy controls are also presented. In benign cases, LRIG1 was overexpressed throughout the layers (A–C). In case 3, koilocytes seemed to be negative for LRIG1 (C). On the other hand, it was generally negative in malignant cases (G–I). Only a small number of LRIG1 positive cells were found in the very surface layer of malignant cases (G–I). On the contrary, EGFR expression was faint in benign cases (D–F), and inversely very strong in malignant cases (J–L). Scale bars = 100 µm.</p

Patient Demographics and Clinicopathological Features.

<p>CIS = carcinoma in situ; SCC = squamous cell carcinoma.</p

Histopathology of 6 OSSN cases.

<p>Histopathology of 6 cases with lower magnification (A–F) and higher magnification (A′–F′). (A) Case 1. Thick, acanthotic squamous epithelium with fibrovascular cores. Some cells had more hyperchromatic nuclei, suggesting a mild degree of dysplasia (arrows in A′). (B) Case 2. Almost all the layers were replaced by large squamous cells, but surface layer presented differentiation (arrow in B′). (C) Case 3. The bottom third to half of the epithelium was transformed to small epithelial cells (arrows). Koilocytes were identified in upper layer (C′). Unlike benign cases, the full thickness of the epithelium was replaced by atypical cells in case 4–6 (D–F). (D) Case 4. Multiple epithelial lobules extended into the subepithelial connective tissue (arrows). Cells have atypical appearance, with darker cytoplasm and pleomorphic nuclei (D′). (E) Case 5. The basement side was irregular with increased inflammatory responses in underlying connective tissue. Within the epithelium, there was abnormal keratinization with central foci of acellular keratin (arrows in E′). (F) Case 6. The infiltrating lobules, nests, and cords were extending from the basement line into the underlying connective tissue with lymphocytic infiltrate within the connective tissue (arrows). The surface hyperkeratosis was prominent (arrowheads) simultaneously with intraepithelial hyperkeratosis (arrows in F′). Cell atypia is prominent (F′). Scale bars = 100 µm.</p

The expression pattern of keratins.

<p>The expression pattern of keratins 4 (A–C, M–O), 13 (D–F, P–R, d–f), 12 (G–I, S–U), 15 (J–L, V–X) in the 6 cases; case 1 (A, D, G, J), case 2 (B, E, H, K), case 3 (C, F, I, L), case 4 (M, P, S, V), case 5 (N, Q, T, W), case 6 (O, R, U, X). The expression pattern of these keratins in conjunctivas (a, d, g, j), limbus (b, e, h, k), and corneas (c, f, i, l) in healthy controls are also presented. K4 and K13 were partially positive or totally positive in benign cases (A–F). In the CIS cases (case 4 and 5), strongly positive pattern (M, P, Q) and weak positive pattern (N) were combined. In an invasive SCC case (case 6), K4 and K13 were generally negative (O,R). K12 expression was negative in both benign and malignant cases (G–I, S–U). K15 was overexpressed in all the OSSN tissue (J–L, V–X). The expression pattern was comparatively limited to the basal layer in benign cases (J–L). All three malignant cases showed notably positive staining (V–X). Scale bars = 100 µm.</p

Ki67 expression in OSSN cases.

<p>(A and B) Representative expression pattern of benign cases (A, case 2) and malignant cases (B, case 4). Ki67 positive cells were disseminated both in benign and malignant samples. The expression pattern was comparatively sparse and limited in the basal area in the benign samples (A), while diffuse and dense in malignant samples (B). (C) Ki67 labelling index was significantly (p<0.002) high in the malignant tissue (33.2%) rather than benign tissue (10.9%). Scale bars = 100 µm.</p

A Clinic-based Survey of Clinical Characteristics and Practice Pattern of Dry Eye in Japan

<p><strong>Article full text</strong></p> <p><br> The full text of this article can be found <a href="https://link.springer.com/article/10.1007/s12325-017-0487-x"><b>here</b>.</a><br> <br> <strong>Provide enhanced digital features for this article</strong><br> If you are an author of this publication and would like to provide additional enhanced digital features for your article then please contact <u>[email protected]</u>.<br> <br> The journal offers a range of additional features designed to increase visibility and readership. All features will be thoroughly peer reviewed to ensure the content is of the highest scientific standard and all features are marked as ‘peer reviewed’ to ensure readers are aware that the content has been reviewed to the same level as the articles they are being presented alongside. Moreover, all sponsorship and disclosure information is included to provide complete transparency and adherence to good publication practices. This ensures that however the content is reached the reader has a full understanding of its origin. No fees are charged for hosting additional open access content.<br> <br> Other enhanced features include, but are not limited to:<br> • Slide decks<br> • Videos and animations<br> • Audio abstracts<br> • Audio slides<u></u></p

Associations between Subjective Happiness and Dry Eye Disease: A New Perspective from the Osaka Study

<div><p>Importance</p><p>Dry eye disease has become an important health problem. A lack of concordance between self-reported symptoms and the outcome of dry eye examinations has raised questions about dry eye disease.</p><p>Objective</p><p>To explore the association between subjective happiness and objective and subjective symptoms of dry eye disease.</p><p>Design</p><p>The study adopted a cross-sectional design.</p><p>Setting</p><p>All the employees of a company in Osaka, Japan.</p><p>Participants</p><p>672 Japanese office workers using Visual Display Terminals (age range: 26–64 years).</p><p>Methods</p><p>The dry eye measurement tools included the Schirmer test, conjunctivocorneal staining, the tear film break-up time, as well as the administration of a dry eye symptoms questionnaire. Happiness was measured by the Subjective Happiness Scale.</p><p>Main Outcome Measures</p><p>Dry eye examination parameters, dry eye symptoms questionnaires, and the Subjective Happiness Scale score.</p><p>Results</p><p>Of the 672 workers, 561 (83.5%) completed the questionnaires and examinations. The mean Subjective Happiness Scale score was 4.91 (SD = 1.01). This score was inversely correlated with the dry eye symptom score (r = -0.188, p < 0.001), but was not associated with objective findings which include conjunctivocorneal staining, low Schirmer test score, or low tear film break-up time. The level of subjective happiness was the lowest in the group without objective results, but reported subjective symptoms of dry eyes (p < 0.05).</p><p>Conclusions and Relevance</p><p>There is evidence of the relationship between subjective happiness and self-reported symptoms of dry eyes. Findings of this study revealed a new perspective on dry eye disease, including the potential for innovative treatments of a specific population with dry eye disease.</p></div

Comparison of dry eye symptom scores between objective and subjective classifications.

<p>SD = standard deviation.</p