Heat-map of significantly differentially expressed miRNAs associated with outcome after adjuvant Tamoxifen treatment.

<p>Data is based on the discovery set (p<6.6e-4, FDR 2.5% and variance >0.1). The green symbols above the heat-map indicate samples from patients with no recurrence, whereas the red symbols indicate samples from patients with recurrence. The heat-map is a standardized intensity plot with the intensities ranging from −2 (green) to +2 (red).</p

Association of miR-7 with tumor grade.

<p>Grade 1 vs. 3 and Grade 2 vs. 3: p = 0.01, and Grade 1 vs. 2: p = 0.02). N = 52 patients (Discovery set).</p

Kaplan-Meier plots of the 10 miRNAs identified in the discovery set.

<p>The bold line represents the patients with a good prognosis, whereas the dotted line represents the poor-prognosis patients. A) Probability of recurrence. B) Probability of overall survival.</p

Characteristics of included patients and their breast cancer tumor (N = 152).

a<p>the average and median were calculated only for the tumors defined as positive, i.e. staining was observed in ≥ 10% of tumor cells by immunohistochemistry.</p>b<p>If the actual percentage was not provided, patients were deemed positive if ER staining was observed in ≥ 10% of tumor cells by immunohistochemistry and/or target protein (ER or PgR) was >10 fmol/mg total protein as determined by biochemistry.</p>c<p>the 5 ER- tumors had a PgR status of 90%, 50%, 90%, 80% and IHC+ (i.e. >10%) respectively.</p><p>Abbreviations: R: patients with recurrence. N: patients without recurrence. Disc.: Discovery set. Test#1: Test set#1. Test#2: Test set#2. Avg: average IDC: invasive ductal carcinoma. ILC: invasive lobular carcinoma. TTR: time to recurrence.</p

Bulletin mensuel de statistique / Institut national de la statistique et des études économiques...

septembre 19801980/09 (N9,A31)

Tumor-Preferential Induction of Immune Responses and Epidermal Cell Death in Actinic Keratoses by Ingenol Mebutate

<div><p>The rapid and strong clinical efficacy of the first-in-class, ingenol mebutate, against actinic keratosis (AK) has resulted in its recent approval. We conducted the first comprehensive analysis of the cellular and molecular mode of action of topical ingenol mebutate 0.05% gel in both AK and uninvolved skin of 26 patients in a phase I, single-center, open-label, within-patient comparison. As early as 1 day after application, ingenol mebutate induced profound epidermal cell death, along with a strong infiltrate of CD4⁺ and CD8⁺ T-cells, neutrophils, and macrophages. Endothelial ICAM-1 activation became evident after 2 days. The reaction pattern was significantly more pronounced in AK compared with uninvolved skin, suggesting a tumor-preferential mode of action. Extensive molecular analyses and transcriptomic profiling of mRNAs and microRNAs demonstrated alterations in gene clusters functionally associated with epidermal development, inflammation, innate immunity, and response to wounding. Ingenol mebutate reveals a unique mode of action linking directly to anti-tumoral effects.</p><p><b><i>Trial Registration</i>:</b> ClinicalTrials.gov <a href="https://clinicaltrials.gov/ct2/show/NCT01387711" target="_blank">NCT01387711</a></p></div

Ingenol mebutate 0.05% gel treatment causes rapid infiltration of T-cells, macrophages, and neutrophilic granulocytes.

<p>Biopsy specimens from all patients (<i>n</i> = 26) were subjected to histopathological evaluation. Skin tissues from the two treatment areas and all time points were assessed, and representative images are depicted. The panels depict hematoxylin & eosin staining as well as immunohistochemical staining for CD4⁺ T-lymphocytes, CD8⁺ T-lymphocytes, CD68⁺ macrophages/histiocytes, and myeloperoxidase (MPO⁺) neutrophils as indicated. The scale bar represents 100 μm.</p

Topical treatment with ingenol mebutate 0.05% gel affects expression of gene clusters relevant for inflammatory and wound healing responses, and induces a characteristic microRNA (miRNA) pattern.

<p>The profiles illustrated are heat-maps and 2-way hierarchical clusterings of deregulated genes of <b>(A)</b> the 95 most variable mRNAs, and <b>(B)</b> the 64 most variable miRNAs across samples (<i>n</i> = 24) for the first six patients in the study. Differentially expressed genes and miRNAs were identified by pair-wise comparison of the following groups: actinic keratosis day 0 (AK0) versus uninvolved skin day 0 (US0), AK2 versus AK0, US2 versus US0, and AK2 versus US2. The expression analysis included variance filtering (VAR >0.2), statistical significance test by analysis of variance (<i>P</i> < 0.01), expression cut-off (>2-fold), and the false-discovery-rate was controlled by the Benjamini-Hochberg procedure (q < 0.05). Microarray data can be found in the GEO repository (GSE63107). Arrows indicate mRNA genes and miRNAs that were selected for validation by quantitative polymerase chain reaction. The colors above the heat map indicate: US0 (dark blue, <i>n</i> = 6), US2 (light blue, <i>n</i> = 6), AK0 (green, <i>n</i> = 6), and AK2 (red, <i>n</i> = 6) samples, respectively. The red and green shades on the heatmap represent up- and down-regulated genes, respectively.</p

Inflammatory skin reactions induced by ingenol mebutate 0.05% gel are more pronounced in actinic keratosis (AK)-lesioned areas compared with uninvolved skin.

<p><b>(A)</b> CONSORT study diagram. This was a phase I, single-center, open-label, within-patient comparison trial to explore the biological effects of ingenol mebutate gel applied once daily for 2 consecutive days in patients with actinic keratosis (AK) in a 25-cm² area on the extremities and a 25-cm² area of uninvolved-skin (US) on the inner upper arm. <b>(B)</b> Typical skin reactions during the course of the trial in AK treatment areas of three representative patients at the dorsum of the hand (upper panel) (<i>n</i> = 26) and uninvolved skin of the inner upper arm (lower panel) (<i>n</i> = 26) at day 0 (baseline), day 1 (after one treatment application), and day 2 (after two treatment applications) as well as during follow-up at days 8 and 29. <b>(B)</b> The composite local skin response (LSR) score is the sum of six individual LSR scores including erythema, flaking/scaling, crusting, swelling, vesiculation/pustulation, and erosion/ulceration, which range from 0 to 4, with higher numbers indicating more severe reactions. This was calculated at each study visit for each patient, with a theoretical maximum composite score of 24. Patients were assessed on days 0, 1, 2, 8, and 29. No unexpected signs of local or systemic toxicity were noted. Illustrated are averages of the LSR; error bars indicate standard error of the mean.</p

Ingenol mebutate 0.05% gel activates cutaneous blood vessels and induces epidermal cell death.

<p>Biopsy specimens from both treatment areas of all patients (<i>n</i> = 26) and all time points were assessed immunohistochemically for expression of CD20⁺ B-lymphocytes, ICAM-1 (CD54), and CD1a⁺ cells. Cleaved caspase 3 and TdT-mediated dUTP-biotin nick end labeling (TUNEL) reactivity indicating apoptotic responses were detected in five of these patients (arrows indicate examples of TUNEL positive epidermal cells). The figure depicts representative slides from all stainings as indicated. The scale bar represents 100 μm.</p