Specialised phenotyping

INFRAFRONTIER2020 project - Trans-national Access call for proposals 
April 2019

 

Context and aims of the call

The INFRAFRONTIER mouse clinics offer the examination of mouse mutants using a broad and standardised phenotypic check-up covering key research areas such as behaviour, clinical chemistry, immunology, energy metabolism, and lung function amongst others. If phenotypes are uncovered by the primary assays, mouse mutants can be subjected to more detailed analyses in hypothesis driven and disease focussed phenotyping pipelines.

In this specialised phenotyping call we provide access to a comprehensive panel of phenotyping tests, relying on standardized and customized protocols in key therapeutic areas.

 

Induced secondary phenotyping screen under acute or more chronic inflammatory conditions

BIOMEDCODE          http://www.biomedcode.com/gr/en

BIOMEDCODE provides access to an induced secondary phenotyping screen under acute or more chronic inflammatory conditions. While primary phenotyping of mutant mouse lines can provide useful information on the involvement of genes in physiology, often their role in pathology can be only revealed when studied in a disease context or following a pathogenic trigger.  Such a trigger can be for example inflammation, which has been implicated in a number of pathologic conditions. To uncover gene function in pathologic processes we offer a specialised phenotyping screen under inflammatory conditions. The proposed screen involves the phenotypic analysis of mutant animals combined with one acute and one chronic inflammatory model. The user’s research interests, the nature of the mutated gene and the strain of the mutant animal under investigation will dictate the selection of the most appropriate combination of one acute and one chronic model, to provide the most informative customized output for the user. Through this secondary screening of mutants under inflammatory and disease conditions, the user will be able to uncover the role of their mutant of interest in pathologic conditions.

 

Functional immune-phenotyping screen by mass cytometry

PHENOMIN-CIPHE           http://ciphe.marseille.inserm.fr/en/

The PHENOMIN-CIPHE mouse clinic provides access to its infrastructure and expertise for analysing mouse mutant lines through specialised functional immune-phenotyping by mass cytometry. To characterize the functional impact, on the immune system, of the invalidation or the overexpression of a gene of interest, cellular immune-phenotype profiling is required. This primary immunophenotyping screen, essentially based on an extracellular labelling, aims at quantifying the various cell populations present within a model by monitoring the expression of specific cell surface receptors in hematopoietic cells. This first approach describing cellular heterogeneity does not assess the effector functions of immune cells. To this end, a more functional secondary immune-phenotyping screen is required and offered in this call.

 

Access to the mouse clinics and their specialised phenotyping screens will be granted based on scientific excellence and supports the development and in-depth characterisation of mouse models for investigating gene function and human pathophysiology.  INFRAFRONTIER will provide open access to all characterised disease models and phenotyping data.

 

Induced secondary phenotyping screen under acute or more chronic inflammatory conditions
Induced secondary phenotyping screen under acute or more chronic inflammatory conditions

BIOMEDCODE      http://www.biomedcode.com/gr/en

Biomedcode can support three projects in this TA call. 

Download application form.

 

The generation of mice with mutated genes resulting in gain and loss of function are very useful tools in studying the functions of specific genes. While primary phenotyping of such animals can provide useful information on the involvement of genes in physiology, often their role in pathology can be only revealed when studied in a disease context or following a pathogenic trigger.  Such a trigger can be for example inflammation, which has been implicated in a number of pathologic conditions.

To uncover gene function in pathologic processes we offer an induced secondary phenotyping screen under acute or more chronic inflammatory conditions. The proposed screen involves the phenotypic analysis of mutant animals combined with one acute and one chronic inflammatory model among the ones outlined below. The user’s research interests, the nature of the mutated gene and the strain of the mutant animal under investigation will dictate the selection of the most appropriate combination of one acute and one chronic model, to provide the most informative customized output for the user.

Acute Models of general inflammation will serve for the fast and sensitive detection of the involvement of the mutated gene in inflammatory mechanisms assessed by the response of mutant mice in cytokine secretion and/or cell recruitment processes. Available acute models include: 1) Lipopolysaccharide (LPS) Induced Cytokine cascade, 2) Zymosan Induced cell recruitment, 3) Thioglucolate Induced cell recruitment

Induced Disease Models will serve for the evaluation of the involvement of the mutated genes in a disease setting, assessed by the response of mutant mice in disease development and progression evaluated at the clinical and histopathological level. Available disease models include: 1) Collagen Antibody induced arthritis (CAIA) (*Arthritomab must be provided by the interested party), 2) TNBS-colitis, 3) IMQ-induced psoriasis, 4) MOG-induced Experimental Autoimmune Encephalomyelitis.

Through this secondary screening of mutants under inflammatory and disease conditions, the user will be able to uncover the role of their mutant of interest in pathologic conditions.

 

Access modalities and the INFRAFRONTIER partner providing the service are described in detail in the application form.

Functional immune-phenotyping screen by mass cytometry
Functional immune-phenotyping screen by mass cytometry

PHENOMIN-CIPHE      http://ciphe.marseille.inserm.fr/en/ 

CIPHE can support three projects in this TA call. 

Download application form.

 

Secondary functional immune-phenotyping screen by mass cytometry

To characterize the functional impact, in the immune system, of the invalidation or the overexpression of a gene of interest cellular immune-phenotyping is required. This primary immunophenotyping screen, essentially based on an extracellular labeling, aims at quantifying the various cell populations present within a model by monitoring the expression of specific cell surface receptors in hematopoietic cells. This first approach describing cellular heterogeneity does not assess the effector functions of immune cells. To this end, a more functional secondary immune-phenotyping screen is required.

The effector function capacities of immune cells were traditionally assayed by extensive characterization of cytokine production, the detection of master gene regulator of transcriptional program and more recently intracellular signaling. We propose to investigate a set of these hallmarks (12 cytokines, 7 transcription factors) at the single cell level on 8 different lineages (CD4 and CD8 T cells, B cells, NK cells, DCs, macrophages, neutrophils and monocytes) by mass cytometry in one set of experiment on same mice.

The service offer entails the analysis of 12 individual age-matched (8-10 weeks) mice at basal stage or upon inflammatory conditions or tumor challenge. Animals will be split equally over two experimental groups CTRL (C57BL/6) and MUT (mutant). One tissue (spleen, lymph nodes or tumor) will be collected and immune cells extracted. CD45+ cells will be enriched using magnetic sorting if necessary. All work will be done according to CIPHE standardized procedures. Cytokine production expression will be evaluated upon brief in vitro restimulation (PMA/Ionomycin + Brefeldin A for 4 hours). Remaining cells will be banked for later analysis if necessary (secondary experiments not funded through this call).

 

Access modalities and the INFRAFRONTIER partner providing the service are described in detail in the application form.