Identification of a recurrent STRN/ALK fusion in thyroid carcinomas.

Thyroid carcinoma is the most common endocrine malignant tumor and accounts for 1% of all new malignant diseases. Among all types and subtypes of thyroid cancers that have been described so far, papillary thyroid carcinoma is the most frequent. The standard management treatment of these tumors consists of surgery, followed by radioiodine treatment in case of high risk of relapse. The most aggressive forms are commonly treated by chemotherapy, radiotherapy or experimental drug testing. We recently reported the case of a patient presenting an anaplastic thyroid carcinoma with lung metastases. Fluorescence in situ hybridization analysis allowed us to detect a rearrangement of the anaplastic lymphoma kinase (ALK) gene in both tumors. The patient was treated with crizotinib and presented an excellent drug response. We present here the subsequent investigations carried out to further characterize this genetic alteration and to assess the prevalence of ALK rearrangements in thyroid lesions. High resolution array-comparative genomic hybridization data complemented by RT-PCR and sequencing analyses, allowed us to demonstrate the presence of a STRN/ALK fusion. The STRN/ALK transcript consisted of the fusion between exon 3 of STRN and exon 20 of ALK. Subsequent screening of 75 various thyroid tumors by RT-PCR revealed that 2 out of 29 papillary thyroid carcinomas exhibited the same fusion transcript. None was detected in other types of malignant or benign thyroid lesions analyzed. These findings could pave the way for the development of new targeted therapeutic strategies in the treatment of papillary thyroid carcinomas and point to ALK inhibitors as promising agents that merit rapid evaluation

Efficacy of antiandrogens in androgen receptor-positive triple-negative metastatic breast cancer: Real-life data

Antiandrogens (AA) have been tested in clinical trials in androgen receptor (AR) + triple-negative breast cancer (TNBC). We aim to assess the clinical benefit rate (CBR) of AA in real life.The primary end-point was CBR at 6 months. Twenty-four patients were assessable and received: abiraterone acetate (62 %), enzalutamide (8 %) and bicalutamide (30 %). CBR at 6 months was 29 % (7/24) with 2 CR, 3 PR and 2 SD. Four patients had a clinical benefit >12 months. Real-life efficacy of AA use in metastatic AR + TNBC are in line with data from published trials

Regulation of senescence escape by TSP1 and CD47 following chemotherapy treatment

Abstract Senescence is a tumor-suppressive mechanism induced by telomere shortening, oncogenes, or chemotherapy treatment. Although it is clear that this suppressive pathway leads to a permanent arrest in primary cells, this might not be the case in cancer cells that have inactivated their suppressive pathways. We have recently shown that subpopulations of cells can escape chemotherapy-mediated senescence and emerge as more transformed cells that induce tumor formation, resist anoikis, and are more invasive. In this study, we characterized this emergence and showed that senescent cells favor tumor growth and metastasis, in vitro and in vivo. Senescence escape was regulated by secreted proteins produced during emergence. Among these, we identified thrombospondin-1 (TSP1), a protein produced by senescent cells that prevented senescence escape. Using SWATH quantitative proteomic analysis, we found that TSP1 can be detected in the serum of patients suffering from triple-negative breast cancer and that its low expression was associated with treatment failure. The results also indicate that senescence escape is explained by the emergence of CD47low cells that express a reduced level of CD47, the TSP1 receptor. The results show that CD47 expression is regulated by p21waf1. The cell cycle inhibitor was sufficient to maintain senescence since its downregulation in senescent cells increased cell emergence. This leads to the upregulation of Myc, which then binds to the CD47 promoter to repress its expression, allowing the generation of CD47low cells that escape the suppressive arrest. Altogether, these results uncovered a new function for TSP1 and CD47 in the control of chemotherapy-mediated senescence

<i>ALK</i>, <i>STRN</i> and <i>STRN/ALK</i> fusion transcript RNA expression and sequencing.

<p>(A) Expression of <i>ALK</i>, <i>STRN</i> and <i>STRN/ALK</i> fusion transcript obtained by RT-PCR in the pulmonary tumor, the thyroid tumor and non tumoral tissue of the initial sample, and in one control sample (C1) are presented. L: molecular weight ladder, bp: base-pair. (B) Expression of <i>STRN/ALK</i> fusion transcript obtained by an <i>ALK</i> specific RT using <i>ALK</i>ex20R1 primer followed by PCR in the same samples. (C) Chromatogram showing the sequence of <i>STRN/ALK</i> fusion transcript at the breakpoint observed in the pulmonary and thyroid carcinomas. The <i>STRN</i> exon 3 (NM_003162) at the 5′ part of the transcript is fused to the <i>ALK</i> exon 20 (NM_004304) at the 3′ portion. (D) Schematic representation of the structure of the fusion transcript, of ALK and STRN proteins and of the predicted fusion protein. For the fusion transcript, initial position of the nucleotides at the fusion are indicated (position on <i>STRN</i> and <i>ALK</i> mRNA). Nu: nucleotide, aa: amino acid, cc: coiled-coil domain. Ensembl genome browser ID: <i>ALK</i> (gene: ENSG00000171094, mRNA: ENST00000389048, prot: ENSP00000373700) <i>STRN</i> (gene: ENSG00000115808, mRNA: ENST00000263918, prot: ENSP00000263918).</p

Genomic profile.

<p>(A) CGH profile obtained from a pulmonary metastasis of the initial case. Genomic alterations are presented and organized on the X axis from chromosome 1 to 22 and X, Y. Log2 ratio values are reported on the Y axis. Significant gains or losses are indicated by blue lines and blue areas above or below each profile, respectively. Chromosome 2 is highlighted with a black box. (B) Enlargements of chromosome 2 and <i>ALK</i> chromosomal region. The chromosome 2 region containing <i>ALK</i> locus is highlighted with a black box. The log2 ratio values of probes covering the proximal regions of <i>ALK</i> and the gene itself are presented. The arrow indicates the breakage region. (C) Enlargements of chromosome 2 and <i>STRN</i> chromosomal region. The chromosome 2 region containing <i>STRN</i> locus is highlighted with a black box. The log2 ratio values of probes covering the proximal regions of <i>STRN</i> and the gene itself are presented. The arrow indicates the breakage region. (D) Schematic representation of potential breakpoints into the two genes at the genomic level (gDNA) according to the CGH data. Agilent CGH probes surrounding the different breakpoints are indicated.</p

ALK Interphase fluorescence <i>in situ</i> hybridization and immunohistochemistry.

<p>(A) On the representative picture of the FISH, realized using the LSI ALK Dual Color Break Apart Rearrangement Probe, we could observe <i>ALK</i> probe break-aparts highlighted by arrows in the two positive PTC cases. Magnification: X1000. (B) Pictures of immunohistochemical labeling for ALK in the two PTC samples. Pictures of hematoxylin, eosin and safran (HES) staining of the two samples are also presented. Magnification for HES: X200, Magnification for IHC: X400.</p

<i>ALK</i>, <i>STRN</i> and <i>STRN/ALK</i> fusion status at mRNA and genomic DNA levels in positive PTC.

<p>(A) Expression profiles of <i>ALK</i>, <i>STRN</i> and <i>STRN/ALK</i> fusion transcript obtained by RT-PCR in two PTC samples and in one control sample (C1) are presented. L: molecular weight ladder, bp: base-pair. (B) Chromatogram showing the sequence of <i>STRN/ALK</i> fusion transcript at the breakpoint observed in the two PTC samples. The <i>STRN</i> exon 3 (NM_003162) at the 5′ part of the transcript is fused to the <i>ALK</i> exon 20 (NM_004304) at the 3′ portion. (C) Products obtained after PCR on genomic DNA using first <i>ALK</i> and <i>STRN</i> forward and reverse primers and then the combination of a <i>STRN</i> forward primer with an <i>ALK</i> reverse primer for case 5 and one control sample (C1). (D) Chromatogram showing the sequence of <i>STRN/ALK</i> fusion at the genomic breakpoint observed in the case 5. One nucleotide at the intronic junction (C) is identical in both fused genes and might be contributed by either of them.</p

Neoadjuvant anthracycline-based (5-FEC) or anthracycline-free (docetaxel/carboplatin) chemotherapy plus trastuzumab and pertuzmab in HER2 + BC patients according to their TOP2A: a multicentre, open-label, non-randomized phase II trial

International audiencePurpose Previous studies have reported the benefit of dual HER2-targeting combined to neoadjuvant chemotherapy in HER2-amplified breast cancer (HER2 + BC). Moreover, besides the cardiac toxicity following their association to Trastuzumab, anthracyclines chemotherapy may not profit all patients. The NeoTOP study was designed to evaluate the complementary action of Trastuzumab and Pertuzumab, and the relevance of an anthracycline-based regimen according to TOP2A amplification status. Methods Open-label, multicentre, phase II study. Eligible patients were aged ≥ 18 with untreated, operable, histologically confirmed HER2 + BC. After centralized review of TOP2A status, TOP2A-amplified (TOP2A+) patients received FEC100 for 3 cycles then 3 cycles of Trastuzumab (8 mg/kg then 6 mg/kg), Pertuzumab (840 mg/kg then 420 mg/kg), and Docetaxel (75mg/m 2 then 100mg/m 2 ). TOP2A-not amplified (TOP2A-) patients received 6 cycles of Docetaxel (75mg/m 2 ) and Carboplatin (target AUC 6 mg/ml/min) plus Trastuzumab and Pertuzumab. Primary endpoint was pathological Complete Response (pCR) using Chevallier’s classification. Secondary endpoints included pCR (Sataloff), Progression-Free Survival (PFS), Overall Survival (OS), and toxicity. Results Out of 74 patients, 41 and 33 were allocated to the TOP2A + and TOP2A- groups respectively. pCR rates (Chevallier) were 74.4% (95%CI: 58.9–85.4) vs. 71.9% (95%CI: 54.6–84.4) in the TOP2A + vs. TOP2A- groups. pCR rates (Sataloff), 5-year PFS and OS were 70.6% (95%CI: 53.8–83.2) vs. 61.5% (95%CI: 42.5–77.6), 82.4% (95%CI: 62.2–93.6) vs. 100% (95%CI: 74.1–100), and 90% (95%CI: 69.8–98.3) vs. 100% (95%CI: 74.1–100). Toxicity profile was consistent with previous reports. Conclusion Our results showed high pCR rates with Trastuzumab and Pertuzumab associated to chemotherapy. They were similar in TOP2A + and TOP2A- groups and the current role of neoadjuvant anthracycline-based chemotherapy remains questioned. Trial registration number NCT02339532 (registered on 14/12/14)

Perioperative Cetuximab with Cisplatin and 5-Fluorouracil in Esogastric Adenocarcinoma: A Phase II Study

Purpose: While perioperative chemotherapy provides a survival benefit over surgery alone in gastric and gastroesophageal junction (G/GEJ) adenocarcinomas, the results need to be improved. This study aimed to evaluate the efficacy and safety of perioperative cetuximab combined with 5-fluorouracil and cisplatin. Patients and Methods: Patients received six cycles of cetuximab, cisplatin, and simplified LV5FU2 before and after surgery. The primary objective was a combined evaluation of the tumor objective response (TOR), assessed by computed tomography, and the absence of major toxicities resulting in discontinuation of neoadjuvant chemotherapy (NCT) (45% and 90%, respectively). Results: From 2011 to 2013, 65 patients were enrolled. From 64 patients evaluable for the primary endpoint, 19 (29.7%) had a morphological TOR and 61 (95.3%) did not stop NCT prematurely due to major toxicity. Sixty patients (92.3%) underwent resection. Sixteen patients (/56 available, 28.5%) had histological responses (Mandard tumor regression grade ≤3). After a median follow-up of 44.5 months, median disease-free and overall survival were 24.4 [95% CI: 16.4-39.4] and 40.3 months [95% CI: 27.5-NA], respectively. Conclusion: Adding cetuximab to the NCT regimen in operable G/GEJ adenocarcinomas is safe, but did not show enough efficacy in the present study to meet the primary endpoint (NCT01360086)