Will Shepheard

Establishment of ALK-autoantibodies, T cell responses against ALK and cell-free DNA as biomarkers and for response monitoring of patients with ALCL

Universitätsklinikum Hamburg-Eppendorf

Supervisor: Wilhelm WOESSMANN, Christine DAMM-WELK


Research objectives:

Children with ALK+ ALCL are currently treated with intensive polychemotherapy with limited efficacy but accompanying acute and long-term toxicity risks. New treatment options for forthcoming studies include targeted agents (ALK TKI, CD30-immunotoxins) and immunostimulatory therapies (vinblastine, PD1-inhibition, vaccination).

Initial Anti-ALK antibody titres as well as minimal disseminated and minimal residual disease (MD/RD) have been described as prognostic factors. However, reliable and validated assays for the detection of anti-ALK antibodies, and the quantification of T cell responses from a limited amount of blood are not available. Furthermore, minimal disease monitoring currently relies on unstable RNA isolated from cells which prohibits measurement in all patients. These limitations hamper further progress in disease monitoring initially and during therapy which is especially important for future personalized therapies.

The goal of the proposed project is:

  1. development and validation of new assays and protocols for disease monitoring of patients with ALCL: First, an assay for quantification of anti-ALK autoantibodies and of anti-ALK T cell responses from small amounts of blood will be developed;
  2. the possibility of using cell-free DNA instead of RNA from cells for minimal residual disease monitoring and detection of secondary aberrations will be evaluated;
  3. DNA-based assays for the measurement of T-cell responses and cell-free (cf)DNA relying on next-generation-sequencing technology will be established.

An ELISA-based assay for the quantification of anti-ALK autoantibodies is envisioned. Assay validation will be performed with existing material collected from ALCL patients. Ultimately, a prospective clinical biomarker trial will be planned, and a protocol developed.

Expected results:

An ELISA-assay for quantification of ALK-auotantibodies will be developed and validated so that it can be distributed to all laboratories in Europe for antibody measurements within clinical studies. An assay for the detection of ALK-specific T lymphocytes from 10ml of whole blood will be established and developed to be used in clinical studies. ctDNA will be measured in plasma and quantified for MRD-monitoring of patients with ALCL.

Planned secondments:

Year 2: Miltenyi Biotech: Development of methods for isolation of immune cell subsets by the provision of a unique training opportunity that will also expose the student to a large biotech company (4 months)

Year 3: Università degli studi di Padova: Access to a different cohort of samples for data validation and training in analysis of small RNA species, techniques that have been built up over many years in the Mussolin lab (3 months)