Thèse Developpement des Cultures des Drgs sur Support Avancés Organoïde-Like pour Ciblage de Douleur Induite par la Chimiothérapie et la Neuropathie Périphérique. H/F - Université de Lille

Établissement : Université de Lille
École doctorale : Biologie Santé de Lille
Laboratoire de recherche : Centre de Recherches en Cancérologie de Lille
Direction de la thèse : George SHAPOVALOV ORCID 0000000197029317
Début de la thèse : 2026-10-01
Date limite de candidature : 2026-04-28T23:59:59

En 2024, le cancer du pancréas était responsable de plus d'un décès sur six, et ce chiffre est en constante augmentation selon les données de l'OMS. Parmi les différentes options thérapeutiques, la chimiothérapie demeure un axe de recherche majeur, mais elle entraîne souvent des effets secondaires graves, notamment des douleurs, ce qui limite son efficacité dans le contrôle de la croissance tumorale. Les agents chimiothérapeutiques (AC), comme l'oxaliplatine, bien qu'efficaces pour freiner la progression du cancer, provoquent fréquemment des effets secondaires sévères tels que l'iléopathie et développement d'une neuropathie périphérique (CIPN, neuropathie périphérique induite par la chimiothérapie). Cette douleur induite par les ACs est liée à la dérégulation des canaux ioniques nociceptifs (CIN), fortement exprimés dans les neurones sensoriels des ganglions de la moelle épinière (DRG, dorsal root ganglion). Les lésions cellulaires induites par les AC sont associées à la libération de chimiokines induisant le stress et à un excès d'espèces réactives de l'oxygène (ROS). De plus, l'activité des CIN est influencée par les ROS et le stress médié par les cytokines, ce qui suggère des voies interconnectées entre la progression du cancer, la chimiothérapie et les réponses douloureuses. Ces facteurs soulignent l'importance du microenvironnement tumoral et neuronal et mettent en évidence la nécessité d'utiliser des complexes cellulaires avancés imitant des tussies, incluant des cellules neuronales et cancéreuses, afin d'étudier de manière fiable les mécanismes de signalisation dérégulés lors de la progression du cancer et après chimiothérapie. Ce projet vise donc à: (1) développer un système organoïde avancé sur une matrice multi-électrodes (puces MEA) permettant de maintenir des neurones DRG isolés dans un environnement proche du milieu naturel grâce à l'incorporation de cellules gliales satellites et de cellules PDAC; et (2) utiliser ce système pour étudier les voies de signalisation impliquant les neurones neuronaux et leur dérégulation par les agents cancéreux.

Mounting evidence relates cancer prognosis with intratumoral neural infiltration, a phenomenon most commonly observed in cancers that arise in highly innervated organs. Thus, while only 30-40% of PDAC patients report abdominal pain at the time of diagnosis, up to 80% of patients develop tumor pain as the disease progresses [1]. Every second patient described cancer pain to be severe. Likewise, the prevalence of pain is estimated to be more than 70 % in the advanced stage of CRC [2]. Most patients report abdominal or back pain [1], while in some patients pain can also occur secondarily, e.g., in case of nerve impingement, duodenal stenosis or metastasis [3]. Tumor innervation may also affect patients' quality of life by causing pain, paresthesia, numbness, and paralysis. Likewise, treatment of PDAC and CRC by commonly used chemotherapy agents (CAs) such as Oxaliplatin, Cisplatin or Paclitaxel, causes severe side effects such as ileopathy and chemo-induced peripheral neuropathy (CIPN) associated with thermal (especially cold) and mechanical allodynia or hyperalgesia in up to 80% of patients [4]. Specifically, CA induced pain sensation is thought to be related to dysregulation of thermal or mechanical (T/M) sensors such as TRPM8, V1, V4, A1 or Piezo2 ion channels, which are highly expressed in dorsal root ganglia (DRG) neurons and inhibitory enteric motor neurons [5]. Multiple recent studies report on changes in expression levels or activity of nocicepting ion channels [6.7], however how such changes are brought about by CA use and, most importantly, how disregulation of these nociceptors is related to CIPN development and associated pain sensation is not well understood.
Among the mechanisms of cellular toxicity of chemotherapy listed above, excessive ROS production plays a significant role as a factor leading to the development of peripheral neuropathy [8]. Similarly, the release of cytokines by chemotherapy-stressed tumor cells is known to induce inflammation and dysregulation of autophagy in DRG neurons via pathways involving TNF- and interleukins [9]. On the other hand, it is also known that the activity of ion channels acting as environmental sensors of painful stimuli is modulated by ROS- or cytokine-induced stress, thus providing an important link between chemotherapy and the development of peripheral neuropathy [10]. Furthermore, our preliminary data shows direct, but not acute, effect of Oxaliplatin on one of the nociceptors proposed to be involved in CA-induced pain and CIPN development, TRPM8. These data strongly support the involvement of intracellular pathways regulating nociceptor activity, and involving various kinases, such as PLC, PKA/C and lipid oxidation in the mechanisms of CA-induced pain, potentially addressing both long and short-term aspects of CA-induced pain, described above.
Collectively, these observations suggest the existence of common pathways promoting enhanced pain sensation following chemotherapy, as well as during pancreatic and colon cancer progression. These common mechanisms are likely to involve inflammation, mitochondrial dysfunction of sensory neurons as a consequence of an excessive production of ROS and cytokines in association with alteration of cytosolic Ca2+ signals linked to enhanced activity or expression of NICs. However how these pathways affect nociceptor ion channels and how their disregulation, in turn, affects the resulting pain is not well understood.This project is dedicated to (1) developing and advanced, organoid-based system on a multi-electrod array (MEA chips) that would allow to support isolated DRG neurons in environment approaching natural by incorporating glia satellite cells and incorporating PDAC cells and (2) use this system to study signaling pathways involving NICs and their dysregulation by CAs. Prior experience of our primary PhD candidate working on organoids is an important aspect of this funding application.

To address the issues described above we propose to develop and advanced, organoid-based system on a multi-electrode array (MEA chips) that would allow to support isolated DRG neurons in environment approaching natural by incorporating glia satellite cells and incorporating PDAC cells and use this system to study signaling pathways involving T/M sensors and their dysregulation by CAs. Using isolated DRG neurons, cultured on an astrocyte support layer incorporating also cells originating from cancer cell lines or tumor tissue we aim to create tissue-like constructs (Neurons on Multisupport - NoM) that would retain:
- Differentiated phenotypes of seed DRG neurons;
- Necessary support microenvironment, via exchange of important factors;
- Introduction of cancer cells, mimicking tumor invasion and corresponding microenvironment changes;
- Access to components of the structure for on-line characterization via electrophysiology and confocal microscopy using immunofluorescent markers, or those sensitive to [Ca2+] or other physiological markers.
Prior experience of our primary PhD candidate, described below, working on organoids is an important aspect of this funding application that will allow this project to progress quickly. The project is organized in 3 task, described in detail below.

Le(a) candidat(e) devra posséder des connaissances de base dans le domaine des canaux ioniques et de la signalisation calcique et sodique. Des connaissances sur les principales techniques biochimiques (culture cellulaire, PCR et qRT-PCR, western blot) et travaux avec cultures cellulaires avancées, organoides sont indispensables; des connaissances de base sur l'imagerie calcique et electrophysiologiques (patch clamp, whole cell and single channel recordings and data analysis) serait souhaitable.