Composite biomarkers defined by multiparametric immunofluorescence analysis identify ALK-positive adenocarcinoma as a potential target for immunotherapy

Roussel, H.; Guillebon, Éléonore De; Biard, Lucie; Mandavit, Marion; Gibault, Laure; Fabre, Elizabeth; Antoine, Martine; Hofman, Paul; Beau‐Faller, Michèle; Blons, Hélène; Danel, Claire; Barthes, F. Le Pimpec; Gey, Alain; Granier, Clémence; Wislez, Marie; Laurent‐Puig, Pierre; Oudard, Stéphane; Bruneval, Patrick; Badoual, Cécile; Cadranel, Jacques; Tartour, Éric

doi:10.1080/2162402x.2017.1286437

Cited by 26 publications

(18 citation statements)

References 48 publications

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“…It can semi quantitatively assess the mRNA signal in target cells with conventional light microscopy [ 88 ]. Finally, multiplex IHC analyses are also being developed to analyze in parallel the ALK status and the expression of certain checkpoint inhibitors, and these analyses can participate in studies into novel therapeutic strategies [ 89 ].…”

Section: Methods For Detection Of An Alk Rearramentioning

confidence: 99%

ALK in Non-Small Cell Lung Cancer (NSCLC) Pathobiology, Epidemiology, Detection from Tumor Tissue and Algorithm Diagnosis in a Daily Practice

Hofman

2017

Cancers

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Patients with advanced-stage non-small cell lung carcinoma (NSCLC) harboring an ALK rearrangement, detected from a tissue sample, can benefit from targeted ALK inhibitor treatment. Several increasingly effective ALK inhibitors are now available for treatment of patients. However, despite an initial favorable response to treatment, in most cases relapse or progression occurs due to resistance mechanisms mainly caused by mutations in the tyrosine kinase domain of ALK. The detection of an ALK rearrangement is pivotal and can be done using different methods, which have variable sensitivity and specificity depending, in particular, on the quality and quantity of the patient’s sample. This review will first highlight briefly some information regarding the pathobiology of an ALK rearrangement and the epidemiology of patients harboring this genomic alteration. The different methods used to detect an ALK rearrangement as well as their advantages and disadvantages will then be examined and algorithms proposed for detection in daily routine practice.

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Section: Methods For Detection Of An Alk Rearramentioning

confidence: 99%

ALK in Non-Small Cell Lung Cancer (NSCLC) Pathobiology, Epidemiology, Detection from Tumor Tissue and Algorithm Diagnosis in a Daily Practice

Hofman

2017

Cancers

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“…64 which cells are targeted by antagonists of inhibitory receptors CD8 + T cells The current dogma supported by various preclinical and clinical data argues in favour of a major role of T cells and especially CD8 + T cells to explain the therapeutic activity of antagonists of inhibitory receptors (figure 2). 65 66 The presence of pre-existing T cells before therapy correlated with the clinical activity of PD-1 and CTLA-4 blockade, as TIL density in melanoma patients treated by anti-CTLA-4 correlated with good clinical response. 67 Another study reported that pre-existing CD8 + T cells at the invasive tumour margin were a prerequisite for the efficacy of PD-1 blockade in a cohort of 15 patients treated for melanoma.…”

Section: Blockade Of Intrinsic Tumour Signalling Mediated By Pd-l1mentioning

confidence: 99%

Mechanisms of action and rationale for the use of checkpoint inhibitors in cancer

et al. 2017

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The large family of costimulatory molecules plays a crucial role in regulation of the immune response. These molecules modulate TCR signalling via phosphorylation cascades. Some of the coinhibitory members of this family, such as PD-1 and CTLA-4, already constitute approved targets in cancer therapy and, since 2011, have opened a new area of antitumour immunotherapy. Many antibodies targeting other inhibitory receptors (Tim-3, VISTA, Lag-3 and so on) or activating costimulatory molecules (OX40, GITR and so on) are under evaluation. These antibodies have multiple mechanisms of action. At the cellular level, these antibodies restore the activation signalling pathway and reprogram T cell metabolism. Tumour cells become resistant to apoptosis when an intracellular PD-L1 signalling is blocked. CD8+ T cells are considered to be the main effectors of the blockade of inhibitory receptors. Certain CD8+ T cell subsets, such as non-hyperexhausted (CD28+, T-bethigh, PD-1int), follicular-like (CXCR-5+) or resident memory CD8+ T cells, are more prone to be reactivated by anti-PD-1/PD-L1 monoclonal antibody (mAb). In the future, the challenge will be to rationally combine drugs able to make the tumour microenvironment more permissive to immunotherapy in order to potentiate its clinical activity.

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“…A multi-parametric immunofluorescence study including ALK, CD8, PD-1 and PD-L1 was recently performed. In comparison to EGFR-mutated or WT lung adenocarcinomas, ALK-positive tumors had a higher expression of PD-L1 and a higher number of intra-tumoral CD8+ T cells orPD-1+CD8+Tcells [19].…”

Section: Discussionmentioning

confidence: 89%

Microfluidics-based immunofluorescence for fast staining of ALK in lung adenocarcinoma

Brajkovic¹,

Procopio²

2018

Preprint

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BACKGROUND Anaplastic lymphoma kinase (ALK) is a key oncogenic driver in lung adenocarcinoma patients and its fusion proteins are routinely assessed. The microfluidic tissue processor (MTP) device is based on a chip-confined low-volume technology allowing for rapid immunohistochemistry/immunofluorescence (IHC/IF) stainings of formalin-fixed paraffin-embedded (FFPE) or frozen tissue samples. METHODS A novel ALK IF protocol was developed for the MTP device using the primary mouse anti-human ALK antibody clone 5A4. FFPE tumor whole sections from 14 resected lung adenocarcinoma patients documented to be ALK positive (ALK+) by automated chromogenic IHC and/or FISH were used. MTP-derived IF immunoreactivity was measured by computerized analysis of digitalized images on individual frames of tumor epithelia and surrounding stroma, using an ImageJ plug-in. RESULTS The 5A4 antibody yielded saturated immunoreactivity at an incubation time of 4 min on a titration curve ranging from 2 to 32 min. Total staining time on the MTP device was 18 min including secondary IgG Alexa Fluor 647. MTP-based ALK IF confirmed all 12 cases; with epithelial signal above stromal staining based on computerized pixel-based measurement. MTP-IF (mean intensity levels 458 to 1301) and chromogenic IHC (H-score 120 to 300) showed an equal range of variation of 2.8 and 2.5 folds, respectively, and a trend for direct correlation (p-value 0.051). CONCLUSION The newly developed protocol for immunofluorescent detection of ALK protein with the MTP device confirms chromogenic IHC results on FFPE lung adenocarcinoma specimens. MTP-based IF is fast and reliable. We foresee this study to be a first step opening the road for further realization of microfluidic-based assays for rapid simultaneous detection of targetable oncogenic and immune-system related markers in their topographical context to investigate tumour heterogeneity and micro-environmental interactions.

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Composite biomarkers defined by multiparametric immunofluorescence analysis identify ALK-positive adenocarcinoma as a potential target for immunotherapy

Cited by 26 publications

References 48 publications

ALK in Non-Small Cell Lung Cancer (NSCLC) Pathobiology, Epidemiology, Detection from Tumor Tissue and Algorithm Diagnosis in a Daily Practice

ALK in Non-Small Cell Lung Cancer (NSCLC) Pathobiology, Epidemiology, Detection from Tumor Tissue and Algorithm Diagnosis in a Daily Practice

Mechanisms of action and rationale for the use of checkpoint inhibitors in cancer

Microfluidics-based immunofluorescence for fast staining of ALK in lung adenocarcinoma

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