US-12624395-B2 - Process for identifying a stress state and/or for assessing the stress response level in a subject

US12624395B2US 12624395 B2US12624395 B2US 12624395B2US-12624395-B2

Abstract

A process for identifying a stress state in a subject, for assessing the stress response level in a subject, for predicting the efficacy of an intervention solution in a subject, for monitoring the efficacy of an intervention solution in a subject and/or for identifying an intervention solution for a subject, having a step of detecting the expression and/or quantifying the expression level, in a sample of the subject, of at least four genes selected from a group made of twenty-six genes, as well as a kit having a way for amplifying and/or detecting the expression of the genes, and uses thereof.

Inventors

Alain Lescure
Luc THOMES
Yves Mercier
Mickaël BRIENS

Assignees

ADISSEO FRANCE S.A.S.
CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE

Dates

Publication Date: 20260512
Application Date: 20200313
Priority Date: 20190315

Claims (6)

1 . An in vitro process for identifying the presence or absence of a stress state in a subject being submitted to a challenge and/or for assessing the stress response level in a subject being submitted to a challenge comprising (a) detecting the expression and/or quantifying the expression level, in a sample of said subject, of at least four genes selected from the group consisting of Anxa1, Anxa2, Chac1, and Postn; and (b) administering to the subject at least one intervention solution to reduce or eliminate the stress state and/or stress response level in the subject, wherein the subject and the challenge are selected from: chickens subjected to heat stress, pigs subjected to heat and inflammatory stress, chickens subjected to a nutritional challenge, and mice subjected to a physical exercise challenge.
2 . The in vitro process according to claim 1 , wherein said sample is selected from the group consisting of muscle tissue, breast tissue, liver tissue, adipose tissue, skin, lymphoid tissue, placental tissue, gastrointestinal tract tissue, genital tract tissue, central nervous system tissue, spinal cord, ganglion of the trigeminal nerve, urine, feces, feathers, tears, sperm, seminal fluid, cerebrospinal fluid, expectorations, bronchoalveolar lavage fluid, gastric secretions, saliva, serum, plasma and blood.
3 . The in vitro process according to claim 1 , wherein gene expression is detected and/or quantified at the mRNA level.
4 . The in vitro process according to claim 1 , wherein gene expression is detected and/or quantified at the protein level.
5 . The in vitro process according to claim 4 , wherein gene expression is detected and/or quantified by ELISA, Western blot, immunohistochemistry, flow cytometry or proteomics.
6 . The in vitro process according to claim 1 , further comprising comparing gene expression levels in said sample of said subject with a reference value or with gene expression levels in a reference sample.

Description

FIELD OF THE INVENTION The present invention relates to a process for identifying a stress state in a subject, for assessing the stress response level in a subject, for predicting the efficacy of an intervention solution in a subject, for monitoring the efficacy of an intervention solution in a subject and/or for identifying an intervention solution for a subject, comprising a step of detecting the expression and/or quantifying the expression level, in a sample of said subject, of at least four genes selected from a group consisting of twenty-six genes, as well as a kit comprising means for amplifying and/or detecting the expression of said genes, and uses thereof. BACKGROUND OF THE INVENTION Stress is commonly defined, as proposed by Hans Selye in its 1956' seminal book “The stress of life” (Edition: McGraw-Hill Book Company), as “a state manifested by a specific syndrome, which consists of all non-specifically induced changes within a biological system”. In particular, stress is a physiological state, wherein individuals exposed to modifications of biotic or abiotic factors, called stressors or challenges, in the environment initiate a response to maintain homeostasis. This stress response will first induce behavioral, biological and physical changes in these individuals, that can ultimately result in tolerance and adaptation. In case adaptation is not completed or fails, the tolerance threshold may be exceeded, resulting in altered biological functions. The inability of an individual to withstand stress or to cope with environmental challenges may lead to many unfavorable consequences, ranging from discomfort to death. In particular, in animal production, a wide range of abiotic stressors has been identified, such as social interactions or rough handling, common farm practices (e.g. castration, dehorning, teeth clipping, shoeing, weaning crowding), improper feeding, exposure to adverse climatic conditions, exercise, work and transport. Stressor exposure occurring during the rearing period is likely to influence the preservation of animal well-being, productivity and performance. Animal growth parameters (such as weight gain, feed intake, Feed Conversion Ratio, mortality and morbidity) are traditionally used to characterize animal performance, but they do not specifically reflect animal stress state. It is therefore essential to be able to identify stress state in a subject (human or animal), through the use of sensitive and robust diagnostic tools. Some stress biomarkers have previously been proposed among circulating hormones (e.g. cortisol, ACTH, adrenalin, ocytocine, vasopressin), catabolism products, such as accumulation of oxidated lipids (e.g. malondialdehyde, isoprostanes, hydroperoxides, oxydated LDL, hexanoyl-lysine) or accumulation of oxidated proteins (e.g. nitrotyrosine, carbonylated proteins), inflammation biomarkers (e.g. hydrogen peroxide, myeloperoxidase), biomarkers of oxidated DNA (e.g. 8-hydroxy-2′-deoxyguanosine), or the measure of the activity of intracellular antioxidant systems (e.g. superoxide dismutase, peroxiredoxines, thioredoxines, glutaredoxines, glutathion peroxidases, glutathion). On this basis, kits have also been developed, such as the Oxyscale diagnostic kit (as described in the patent EP2017623) or the QIAGEN Oxidative stress RT2 profiler PCR arrays, which rely on the measure of the expression of a large number of genes. Various genes have also been described as being involved in particular cellular models of stress, but it is well-known to the person skilled in the art that results obtained in vitro are rarely translatable in vivo, and it cannot be deduced from these studies which of these genes, if any, could be useful for identifying a stress state in a subject (human or animal). Overall, all these biomarkers show several limitations, in particular because they are often transient, and specific of the type of stressor and/or the species considered. Indeed, these biomarkers are not easily transferrable between species and/or stress conditions, and no satisfactory universal biomarker has been identified yet. In this context, it was discovered by the inventors, completely unexpectedly, that the detection and/or quantification of the expression of at least four genes selected in a list of twenty-six genes is useful to circumvent these problems. These genes represent biomarkers which are independent of the type of stressors and the species considered. DESCRIPTION OF THE INVENTION Therefore, the present invention relates to an in vitro process for identifying the presence or absence of a stress state in a subject, and/or for assessing (and/or for quantifying) the stress response level in a subject, comprising a step of detecting the expression and/or quantifying the expression level, in a sample of said subject, of at least four genes selected from the group consisting of Ankrd33b, Anxa1, Anxa2, Chac1, Cidea, Col1a1, Col12a1, Col14a1, Efemp1, G0s2, Gfpt2, Hmox1, Kctd12, Ke