PAVE
A nanovaccine Approach for the treatment of Pancreatic Cancer

Work package 1 - PDAC surgery, histology and sample processing in humans, mice and pigs

Objectives: to standardise the protocols and logistics for collecting, storing and transporting of both fresh and bio-
banked human, mouse and pig PDAC samples, for their subsequent immunological analysis later in WP2, WP3 and
WP6 and to perform state-of-the-art pCLE single cell resolution imaging, together with target-specific (CEA, cRGD
and αvβ6) fluorescent probes, to study PDAC cell/stroma interaction during the process of tumour development in mice.

Work package 2  - Identification and high throughput synthesis of antigens specific for PDAC

Objectives: to select immunogenic antigens and epitopes that are expressed in PDAC of humans, mice and pigs for
use in WP5 in our nanovaccine design and to identify PDAC associated antigens in order to provide all the synthetic
antigens for the PAVE project.

Work package 3 - Identify immunome differences in the tumour environment before/after its disruption

Objectives: a) to analyse the immune cell states of the TME in mice and pigs, both, before and after its disruption with
intratumoural hyaluronidase and/or CD40 agonists and b) to perform an analysis of the immune cell states in human
subjects, as a translational reference, both in healthy volunteers and patients

Work package 4 - Transgenic porcine model of PDAC and development of a panel of porcine immunological reagents

Objectives: To a) develop a toolbox of pig-specific immunology reagents and assays, b) create an “oncopig” with
Cre recombinase inducible porcine transgenes encoding PDX1, KRASG12D and TP53R167H and c) produce a T cell
specific reporter system pig to monitor cancer vaccine efficacy. T4

Work package 5 - Provide immunologic and molecular imaging readouts to validate the efficacy of the nanovaccines

Objectives: to develop a suite of “species-specific” assays to measure the response of the immune system appropriately
and according to the species in question. One important task to address is the ability of the nanovaccines, with both
depletion and non-depletion of the TME, to increase the co-stimulatory capacity of DCs in the human, murine and
porcine models.

Work package 6 - Nanovaccine encapsulation of tumour antigens, immunomodulators and imaging contrast agents

Objectives: to test the concept of a multifunctional and multicomponent nanovaccine with two different carrier formats:
a) immunoliposomes and b) Poly Lactic-co-Glycolic Acid (PLGA) NPs. These will be prepared with epitopes of
TAAs encapsulated together with an optimal combination of TLR agonists and/or alpha-GalCer targeted to DCs. Click
chemistry will be applied for introducing the ligands onto the nanovaccine surface. The external coating with specific
ligands of the mouse and human DC receptor (e.g. DEC-205, CD40 or CD11c peptides) should guarantee that these
nanovaccines specifically target DCs in vivo. A study will be performed of receptor interactions and intracellular routing
of modified nanovaccines that target DCs in cancer, in order to improve the efficacy of targeted nanovaccine and to
maximize antigen uptake.