Obtaining and analyzing plasmasamples from patients with non small cell lung cancer, referred for radiation treatment and/or chemotherapy

The main aim of anticancer therapies is to exert cytotoxic effects on cancer
cells. Most contemporary anticancer therapies kill cancer cells through a
non-immunogenic pathway of cell death and are thus unable to *revive* and
mediate anticancer immune responses. However, it recently emerged that some
anticancer therapeutic modalities are capable of inducing a cell death
subroutine called immunogenic cell death (ICD) that can mediate specific,
sustained anticancer immunity (1). These observations have marked the beginning
of intense research into immunoadjuvant or anticancer immunity inducing *side-
effects* associated with anticancer therapies.
No published data about human biomarkers for ICD is available.
Recently it has emerged that ICD may also be associated with a *viral
response-like chemokine signature (VCS)* capable of acting as both *find me*
signal (for granulocytic myeloid cells) and *keep away* signal (for immature
monocytic myeloid cells) - further details of this paradigm are under
investigation (8).
ICD has been found to be associated with several immune effector signatures
(mainly defined as such, for predominantly solid cancers) including positive
dendritic cell (DC) maturation (phenotypic and function) and positive
regulation of T cell immunity (proliferation and Th1 polarization-type cytokine
signature) (10).
The presence of determinants of ICD can be confirmed through the strategy of
following biomarkers (in non-hematological cancers)
We will also investigate serum-associated exosomes as possible biomarkers of an
efficient antitumor response. A secondary endpoint is defined as the cytokine
profile that is linked to a Th1 or Th2-phenotype. A third endpoint consists of
detecting signs of cardiovascular damage with known myocardial markers and
vascular markers. This endpoint is tied to the effects of radiation therapy on
the cardiovascular system. We need to stress that the mentioned lists of
potential biomarkers is non-exhaustive, as this rapidly evolving field might
steer us into a new direction while the study is ongoing. We will keep up with
literature and will implement new biomarker targets into our panel as required.

The aim of this pilot study is to investigate the hypothesis that certain
biomarkers of ICD that were identified in vitro or ex vivo are detectable in
patient sera following radiotherapy and/or chemotherapy. Radiotherapy alone or
concurrent cisplatin-doublet and radiotherapy will be investigated. We will
conduct this pilot study to gather initial data to build upon in future
clinical trials, as there is no in vivo data available on this topic.

40 patients will have a bloodwithdrawal of 25 ml at three points during their
treatment:


For concurrent CT/RT
- Immediately before the first fraction of RT
- Immediately before the third fraction of RT (commonly 48h)
- Immediately after the last or second-to-last fraction of RT


For RT only:
- Immediately before the fraction of RT
- Immediately before the second fraction of RT (commonly 48h)
- Immediately after the last fraction of RT

3 blood withdrawals of 25ml

The burden consists of 3 extra blood withdrawals of 25 ml, with the minimal
risk of damage to a vein or nerve.