WG 2 Mye-SOP

Photo: © German Biobank Node

WG 2 focuses on the validation and standardisation of existing myeloid-related biomarkers. These biomarkers are extracted from literature mining utilising the expertise of Action members also active in WG 1. Those candidates are thus mostly derived from wet lab experiments, but (i) have not yet been properly validated due to a lack of adequate tools for their detection, or (ii) have only been validated in one or a limited number of diseases. Hence, in the first instance WG 2 is establishing the standard operating procedures (SOPs) for the evaluation of existing biomarkers, and, as WG 1 gradually makes progress, intensifies its interaction with WG 1 to explore additional novel biomarkers. WG 2 finally tests the principal applicability of these signatures and markers on real-life clinical samples and, for this purpose, collaborates with WG 3.

Working Group 2 has the following specific tasks:

  1. Existing, but as yet not fully explored signatures and biomarkers should be validated at the protein level on circulating cells or in tissues of various DACI, when applicable. Detecting protein expression will in the first instance require an assessment of the (commercially) available antibodies, their use in published literature and, if needed, the generation of novel antibodies (eventually in collaboration with companies).
  2. Novel antibody panels will be designed for the detection of DACI-associated myeloid cells via appropriate technology such as multi-colour flow cytometry, multiplexed immunohistochemistry or FISH. These panels will undergo a first validation through exploratory staining on material from patients with DACI.
  3. Constitute a task force of groups willing and capable of pre-testing potential biomarker panels on an exploratory level.
  4. Develop standardised SOP-like protocols for myeloid-associated biomarkers.
  5. Train ECI Action members with a special interest in translational biomarker research to conduct biomarker staining and perform analysis.
  6. Translate SOP-like protocols onto archived tissue material obtained from patients with DACI.

These tasks will be achieved through the following activities and networking tools:

  1. WG 2 members will have frequent meetings, in person or online, with WG 1 members to discuss which biomarkers and signatures are promising, starting with existing biomarkers but gradually moving to newly discovered biomarkers. Such markers should be withheld for the subsequent testing in wet-labs. This includes important decisions on whether similar biomarkers can be used for various DACI, or whether disease-specific biomarkers/signatures will be needed.
  2. WG 2 will identify companies with whom to collaborate for the development of antibodies not presently available on the market. Companies will be selected based on their core technological expertise (generation of tools, methods, technologies).
  3. WG 2 will organise workshops with individual companies to assess the company’s suitability and willingness to collaborate on this project and to generate an antibody suitable for immunostaining.
  4. If possible, the same samples will be shared between the different WG 2 laboratories to independently evaluate the suitability of the selected biomarkers. Markers will then be discussed with WG 3 members to relate these markers to clinical information and to determine their suitability for testing on biobanked material.
  5. A smaller number (3-5) of expert labs will join forces to develop SOP-like protocols by exchanging and testing protocols, by web-conferencing and by engaging in STSMs.
  6. A training school on biomarker analysis in tissues and digital pathology will be Companies providing (hardware or software) products in this field will be invited to participate and present their solutions; also as a means of reaching out to private sector stakeholders. STSMs will be organised to train ECIs in SOP application. It is envisioned that 50% of the ECIs will be recruited from ITC countries.

Action members trained under task 6 will apply biomarker candidates via SOP-like protocols on DACI tissues in collaboration with clinical and biobank experts from WG 3.

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