ΕΣΠΑ 2021-2027
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Τhe Institute of Chemical Biology (ICB) employs a set of different biomarkers linked to a variety of human diseases (including cancer and neurodegenerative diseases) and ageing. Analyses are accompanied by detailed service contracts between ICB and the interested parties, defining the selected biomarkers to be tested, the number of samples, the time schedule and the budget. The biomarkers employed include:

Ageing-Related Biomarkers

1. Advanced Glycation End products (AGEs)

Advanced Glycation End products (AGEs) accumulate over lifetime and the accumulation is further enhanced with various conditions (e.g., enhanced oxidative stress, diabetes mellitus, renal failure and cardiovascular disease). Levels of AGEs are determined in serum, plasma and other biological fluids through ELISA.

2. Apolipoprotein –J / Human Clusterin

The different functions of apolipoprotein J (ApoJ)/clusterin (CLU) and its pivotal action in the regulation of life expectancy and progression of age-related diseases have emerged CLU as an excellent biomarker of cellular senescence and organismal ageing. CLU levels are assessed in various sample types (e.g. plasma, cell culture extracts and supernatants) through ELISA.

3. Epigenetic modifications – Global methylation profiling

It is well established that cellular and organismal aging is accompanied by changes in gene expression DNA methylation patterns that, in turn, alter gene expression. Analysis of epigenetic modifications and the potential reversal through treatment with specific compounds is performed on isolated DNA (extracted from whole blood or cell culture) and the results are produced in comparison to existing standard reference curves.

4. Oxidized Proteins levels

Accumulation of proteins undergoing oxidative modifications is enhanced during the progression of ageing, cellular senescence and diseases linked to increased oxidative stress. Levels of oxidized proteins are determined through ELISA, measuring the carbonylated groups on proteins.

5. Proteasome content and activity

The proteasome is the main proteolytic cellular system, responsible for the degradation of normal and damaged, in any way, proteins. Proteasome quantity and activity decline with ageing as well as in various age-related diseases. These levels can be positively affected by implementation of comprehensive lifestyle interventions, e.g., nutrition, reversal of nutritional deficiencies, proper exercise and administration of natural, bioactive compounds. Proteasome levels are determined through ELISA and proteasome activity is detected in crude protein extracts by measuring fluorescence emission following the hydrolysis of the fluorogenic peptide LLVY-AMC.

6. Stem cells physiology

Stem cell exhaustion is one of the 12 fundamental hallmarks of ageing, causally linked to ageing and age-related diseases. Human adult mesenchymal stem cells (hMSCs) are considered to be very promising tools for several cell-based therapies. Nevertheless, adult SCs from elderly donors, who are actually the ones requiring most frequently tissue repairing interventions, have a compromised functionality as a result of ageing. A variety of tests is performed to assess pluripotency, differentiation potential and proliferation capacity of adult stem cells, derived from different sources.

7. Telomere length measurement

Telomeres, the heterochromatic structures at the chromosomal ends, are gradually decreased with cellular senescence and telomere shortening has been associated with various chronic diseases. Relative telomere length is assessed by monochrome multiplex quantitative PCR in isolated genomic DNA from Peripheral Blood Mononuclear Cells (PBMCs).

8. Tyrosinase Activity Assay

Hyperpigmentation is a cosmetically important condition, seen most often in middle-aged and elderly individuals, resulting from exposure to ultraviolet light, certain drugs or chemicals, hormonal alterations or the existence of certain diseases. Tyrosinase is the main copper-containing enzyme, catalyzing several steps in the melanin pigment biosynthetic pathway, thus considered mainly responsible for the age spots. Several compounds can exert whitening effects via topical application. The whitening properties, including the efficacy and optimal dosing of candidate de-pigmenting agents are tested in B10F16 melanocytes, where tyrosinase levels are measured and melanin is quantified using a photography-based method.

Toxicity Testing

Τhe Institute of Chemical Biology (ICB) also employs the nematode Caenorhabditis elegans as a model for toxicity testing of natural or synthetic compounds, extracts and nano-formulations. Analyses are accompanied by detailed service contracts between ICB and the interested parties, defining the number of samples, the time schedule and the budget. The analysis include:

1. Toxicity testing in the nematode Caenorhabditis elegans

Physiological characteristics such as pharyngeal pumping, defecation rate and motility of the nematode Caenorhabditis elegans are highly affected by compounds and environmental factors, thus representing widely accepted toxicity markers, respecting the 3Rs (Replacement, Reduction, and Refinement). They are assessed in the presence of various concentrations of the compounds to be tested.

(CONTACT: Dr. Niki Chondrogianni, [email protected], Dr. Stathis Gonos, [email protected])

2. ADME-Tox (IN3©)

Pharmacokinetic properties, Absorption, Distribution, Metabolism, Excretion (ADME), and the toxicity of xenobiotics are critical factors in decision-making for pharmaceutical R&D and planetary health challenges. Our IN3 framework offers comprehensive ADME-Tox services (B2B, B2C) spanning in silico, in vitro, and in vivo approaches, backed by long-standing expertise in the field. FYI: ADME-Tox

(CONTACT: Dr. Theodora Katsila, [email protected])

 

Apart from service contracts with interested parties, there is also the option of research service/study contracts, where ICB researchers, highly renowned for their expertise in the ageing field, are involved in the experimental design, the results interpretation and the overall project planning and implementation. Detailed analysis of potential anti-ageing and healthspan properties of natural or synthetic compounds or extracts may be performed. The study is accompanied by detailed research service/study contracts between ICB and the interested parties, defining the selected analyses to be performed, the number of compounds to be tested, the time schedule, the budget and other necessary arrangements. In this context, analyses that may be performed include:

Analyses in Human Primary Cells

1. Cellular senescence

Cellular senescence, an irreversible cell cycle arrest, is 1 of the 12 fundamental hallmarks of ageing, and is considered as causal in ageing and an important modulator for an ever-growing number of diseases of late life. Delay of cellular senescence has emerged as a potential route to keep people healthier for longer. Cellular senescence is assessed through the evaluation of the replicative lifespan of human primary fibroblasts (total number of cumulative population doublings) in the presence of compounds to be tested compared to the control cultures.

2. Biochemical biomarkers of cellular senescence

Several cell cycle regulators, have been shown to increase along with the progression of senescence, such as p16 and p21. Simultaneously, the accumulation of these markers is delayed upon anti-ageing remedies. The protein expression levels of these specific cell cycle regulators are revealed through immunoblotting at different time points throughout the replicative lifespan of human primary fibroblasts in the presence of compounds to be tested and the relative control cultures.

3. Senescence-associated β-Galactosidase staining (SA-β-galactosidase activity)

SA-β-galactosidase activity is performed at different time points throughout the replicative lifespan of human primary fibroblasts in the presence of compounds to be tested compared to the control cultures.

Analyses in the Nematode Caenorhabditis Elegans

1. Healthspan assays

Physiological characteristics of the nematode Caenorhabditis elegans, such as pharyngeal pumping, defecation rate and motility of the nematodes are highly affected upon the progression of ageing. Enhancement of healthspan is accompanied by a delay of the alterations of these characteristics. Healthspan properties are assessed in the presence of compounds to be tested in test and control populations at different timepoints of the nematode’s lifespan.

2. Lifespan assays

The nematode Caenorhabditis elegans is an ideal model to study the anti-ageing properties of compounds or extracts in the organismal level. Although standing early in evolution, most of the pathways implicated in human ageing are conserved in the nematode, thus emerging C. elegans as a highly valuable organismal model in the ageing field. On top of that, C. elegans, as an invertebrate, respect the framework for ethical animal use, known as the 3Rs: Replacement, Reduction, and Refinement. Lifespan assays are performed in the presence of compounds to be tested and the median and maximum lifespan in test and control populations are revealed.

(CONTACT: Dr. Niki Chondrogianni, [email protected], Dr. Stathis Gonos, [email protected])

 

 

 

Τhe Institute of Chemical Biology (ICB) employs different biomarkers linked to a variety of human diseases (including cancer and neurodegenerative diseases) and ageing. Analyses are accompanied by detailed service contracts between ICB and the interested parties, defining the selected biomarkers to be tested, the number of samples, the time schedule and the budget. The biomarkers employed include:

Cancer-Related Biomarkers

Individual groups of ICB have interest/activities in Clinical and Translational research projects in colon cancer, lung cancer, breast cancer, pancreatic, prostate cancer, glioblastoma, melanoma..

The predictive/prognostic tumour biomarkers/technologies in sporadic, solid cancers for precision oncology include:

ADME-Tox in cancer

Pharmacokinetic properties, Absorption, Distribution, Metabolism, Excretion (ADME), and the toxicity of xenobiotics are critical factors in decision-making for pharmaceutical R&D and planetary health challenges. Our IN3 framework offers comprehensive ADME-Tox services (B2B, B2C) spanning in silico, in vitro, and in vivo approaches, backed by long-standing expertise in the field. FYI: ADME-Tox

Apart from service contracts with interested parties, there is also the option of research service/study contracts, where ICB researchers, highly renowned for their expertise in the translational research cancer fields, are involved in the experimental design, the results interpretation and the overall project planning and implementation:

  •  Genomic, epigenomic and associated bioinformatic analysis of tumour clinical samples CONTACT Dr Alexandra Voutsina, ([email protected]), Dr Panos Georgiadis ([email protected]), Dr Olga Papadodima ([email protected]), Dr Alex Pintzas ([email protected])
  •  Metabolomic analysis of tumour clinical samples CONTACT Dr Maria Zervou  ([email protected]),  Dr Panos Georgiadis ([email protected])
  •  Multiomic and bioinformatics analysis CONTACT Dr Panos Georgiadis ([email protected]),  Dr Olga Papadodima ([email protected]), Dr Maria Zervou ([email protected]),
  • Predictive biomarkers of drug testing in cancer models CONTACT Dr Dimitra Mitsiou ([email protected]), Dr Alex Pintzas ([email protected]), Dr Panos Georgiadis ([email protected]) Dr Olga Papadodima ([email protected])
  •  Repurposing related biomarkers in cancer (CONTACT: Dr. Theodora Katsila, [email protected])
    1. State-of-the-Art Drug/ Advanced Material Repurposing
      Dry- and wet-lab workflows combining multi-omics, EV profiling, computational biochemistry and AI to prioritize mono- and combination therapies with mechanistic rationale.
    2. IN3 Tri-Modal Framework (in silico · in vitro · in vivo)
      All studies are orchestrated within IN3ã, a tri-modal innovation engine that fuses computational pipelines, experimental assays and translation pathways into a single framework to fast-track advanced materials, smart systems and therapeutics from concept to society.
    3. Extracellular Vesicle (EV) Multi-omics Profiling
      Comprehensive EV analytics from biofluids with emphasis on multi-omics cargo, resulting in candidate EV biomarker panels with performance metrics.
    4. Extracellular Vesicle (EV) Engineering
      Comprehensive EV analytics resulting in fit-for-purpose EV applications with performance metrics.
    5. Inter-individual Variability Mapping upon Xenobiotics Profiling
      Comprehensive analytics resulting in proteoform-based mechanistic insights and fit-for-purpose applications with performance metrics building upon multi-omics datasets.
  • Targeted genetic and epigenetic analysis in clinical samples (tissue and biofluids) CONTACT Dr Alexandra Voutsina, [email protected], Dr Panos Georgiadis ([email protected]), Dr Alex Pintzas ([email protected])
  • Tumour Immune biomarkers CONTACT Dr Panagiota Stamou ([email protected]), Dr Alex Pintzas ([email protected])