DECODING THE IMMUNO-INFLAMMATORY AXIS IN RARE NON-MEDULLARY THYROID CANCER AS AN INNOVATIVE APPROACH FOR NOVEL COMBINATORY THERAPEUTIC APPROACHES

StrategyThe Department of Research, Development and InnovationMedFUTURE Research CentreBiobaseFaculty Research Centres
Biblioteca „Valeriu Bologa”
Editura „Iuliu Haţieganu”
Revista Medicine and Pharmacy Reports

 

Project title: DECODING THE IMMUNO-INFLAMMATORY AXIS IN RARE NON-MEDULLARY THYROID CANCER AS AN INNOVATIVE APPROACH FOR NOVEL COMBINATORY THERAPEUTIC APPROACHES

 

Contract no: 760067/23.05.2023

Code: 81/15.11.2022

Budget: 7.000.000 lei

Project Director: Prof.Dr. ROMANA NETEA MAIER, Radboud UMC Nijmegen, Netherland

 

Summary: The current project investigates the unmet clinical need of patients suffering from poorly differentiated thyroid cancer and anaplastic thyroid cancer resistant to standard therapies with poor outcomes. A major feature of these cancers is the inflammatory pattern that promotes tumor apparition, indicating that tumor inflammatory microenvironment, a new hallmark of cancer, promotes tumor progression and metastasis. The major goal of the proposed research is to comprehensively assess the immune imbalance in aggressive forms of thyroid cancer as an innovative approach to identify personalized combinatoric therapeutic approaches that target immune ecosystem disbalances as a novel strategy to enhance the response to ICI. During the 36 months of this project, an innovative approach will be developed for decoding the dynamics of the immune landscape in PDTC and ATC using high-throughput techniques complemented by in vitro functional assays and in vivo testing of combinatoric therapeutic approaches. This will be realized through several scientific aims, including characterising different immune alterations in aggressive thyroid cancers using high throughput technologies. The impact these alterations could have on how these tumors are unresponsive to conventional therapies will be investigated and validated in both in vitro and in vivo models. The in vivo developed model will allow us to validate new combinatorial therapeutic strategies in clinical practice.

General aim: To comprehensively assess the immune imbalance in aggressive forms of thyroid cancer (PDTC and ATC) as an innovative approach to identify personalized combinatoric therapeutic approaches that target immune ecosystem disbalances as a novel strategy to enhance the response to ICI.

 

Specific Objectives:

SO1. To establish a large integrated research sample repository for aggressive thyroid tumors.

SO2. To characterize the functional alterations of the immune ecosystem in PDTC and ATC (both at systemic and tumor level) using high-throughput techniques.

SO3. To assess the impact of the immune dysregulation using in vitro co-culture models of PDTC and ATC.

SO4. To validate combinatoric treatment strategies in aggressive TC in in vivo models as a first step towards future clinical applications.

 

Expected results:

  • Preparation of the necessary documentation to obtain approval from the Ethics Committee for conducting the research study.
  • Updating standard operating procedures for the collection, anonymization, storage, and processing of biological samples.
  • Creating a biobank of aggressive thyroid cancer (ATC and PDTC) tumor samples, together with an associated database.
  • Developing a specific protocol for the mutational analysis of samples from the biobank (thyroid cancer) using NGS.
  • Obtaining the proteomic profile of tumor samples and integrating them with immunological data and inflammatory markers.
  • Selecting cell lines for co-culture experiments.
  • Developing documentation on the alterations identified following single-cell experiments in PDTC patients.
  • Identifying immunomodulators to be tested in combination with ICI.
  • Developing a report on the validation of combinatorial treatment.
  • Implementation of an effective project management framework, including progress monitoring, publication of results in ISI articles (5 articles), and dissemination through participation in scientific conferences.

 

Results obtained:

  • The ethics documentation was prepared and submitted in accordance with current requirements, and approval was obtained from the Ethics Committee.
  • A set of biological samples and associated database, including biological matrices collected from 27 patients (16 paraffin-embedded tumor tissue samples and 8 goiter samples), as well as a structured clinical dataset for 43 patients diagnosed with various forms of thyroid carcinoma (anaplastic, follicular with poorly differentiated components, insular, mixed medullary and undifferentiated, papillary—classic and poorly differentiated types, poorly differentiated, and undifferentiated carcinoma metastases); database currently being updated until the project’s completion. – Sumar baza de probe biologice
  • Inclusion and exclusion criteria defined and applied for the selection of biological samples.
  • Gene expression dataset obtained through microarray studies, resulting from the comparative analysis of the anaplastic cell lines C643 and Cal-62 versus the normal cell line HPTE, used to identify genetic alterations and select compounds for further testing. Purchased pharmacological screening panel containing 263 compounds (Selechem – L4800 Small Molecule Immuno-Oncology Compound Library), as well as a set of experimental results regarding the evaluation of 27 compounds, of which 7 demonstrated significant inhibitory effects on cell viability (Halofuginone, Fludarabine phosphate, ENMD-2076, Dorsomorphin (Compound C) 2HCl, ITF3756, ENMD-2076 L-(+)-Tartaric acid, PF-477736), enabling the identification of candidates with therapeutic potential and the validation of the cellular pathways investigated.
  • Certificate of professional training obtained by Dr. Raduly Lajos for participating in the course “3D Cell Cultures: From Organoids to Complex and Standardized Human Models, Physiopathological Applications, Cancer,” organized by the National Center for Scientific Research (CNRS) and held at the Lyon Cancer Research Center, Lyon, France.
  • Dataset on the plasma miRNA expression profile obtained through microarray analysis, resulting from the comparative evaluation of a group of 8 patients and 8 healthy volunteers (4 men and 4 women), highlighting changes associated with the investigated pathology.
  • Protein extraction protocol optimized for subsequent analyses.
  • A set of samples prepared for transcriptomic analysis, consisting of total RNA extraction from 46 PDTC and ATC tumor tissue samples, with qualitative and quantitative quantification using Nanodrop, from which 16 samples were selected for Spatial Transcriptomics analysis. – Lista selectie probe pentru ST
  • A well-established method for labeling and characterizing anaplastic thyroid cancer (ATC) cells and macrophages using cell co-cultures and testing therapeutic compounds. –  Tumoroizi pe doua linii celulare – control vs tratament
  • A database of compounds selected for testing, accompanied by a protocol for analyzing cellular drug sensitivity.
  • Biological samples prepared for proteomic analysis using OLINK technology and the REVEAL panel.
  • Co-cultures established between anaplastic thyroid carcinoma cells and THP1 cells, using inserts with a pore size of 0.4 µm, which allow communication between the two cell types exclusively through biological and biochemical mediators.
  • Standardized murine models established for the induction of anaplastic thyroid carcinoma, with animals distributed into distinct groups (control/treatment, immunocompetent/immunodeficient, subjected to immunological stimulation).

 

H2020 Project Applications:

  1. Coordinator in proposal H2020 – European Quality and Innovation in Translational Oncology, Proposal ID 101311162, Acronym EQUITY
  2. Partner in proposal H2020 – HORIZON-HLTH-2025-01 HORIZON-RIA, On-Chip Digital Validation of Allogeneic Cannabinoid Receptor-Based eNK Cell Therapy Targeting Personalised Lung Cancer Niche, Proposal: 101286842, Acronym traiNK-ceLL

 

Dissemination of results:

Participation in international conferences:

 

Participation in national conferences:

  • Human Functional Genomics Romania Conference, September 3-4, 2025, Cluj-Napoca, Romania, poster presentation entitled “ITF3756 Exhibits Inhibitory Effects in Anaplastic Thyroid Carcinoma.” https://hfgpr.com/hfgpr-conference/
  • Zilele Universității de Medicină și Farmacie „Iuliu Hațieganu” Cluj-Napoca (“Iuliu Hațieganu” University of Medicine and Pharmacy Days, Cluj-Napoca), ediția 2025,  8-13 decembrie 2025, https://medpharmareports.com/public/public/Supplements/2025-supplement-1.pdf
    • page 111 –  Ekaterina Isachesku,  ITF3756 as a promising anticancer compound in anaplastic thyroid carcinoma
    • page 130 –  Ecaterina Isacescu,  Identification and functional evaluation of novel therapeutic candidates in anaplastic thyroid carcinoma

 

Publications:

Andreea Bojoga, Pepijn van Houten, Martin Jaeger, Katrin Rabold, Birgitte Walgreen, Liesbeth van Emst, Dumitru Ioachim, Ilse van Engen-van Grunsven, Corin Badiu, Romana T. Netea-Maier, High Local and Systemic Expression of Pentraxin-3 in Anaplastic Thyroid Cancer, Int. J. Mol. Sci. 2025, 26, 11335, https://doi.org/10.3390/ijms262311335

 

The Team

  • Prof. Dr. Romana Netea-Maier, Director Proiect
  • Prof. Univ. Dr. Neagoe Ioana, Cercetător cu experiență
  • Braicu Cornelia, Cercetător cu experiență
  • Pop Laura, Cercetător cu experiență
  • Georgescu Carmen, Cercetător cu experiență
  • Piciu Doina, Cercetător cu experiență
  • Bica Cecilia, Cercetător Post-doctorand
  • Budișan Liviuța, Cercetător Post-doctorand
  • Chira Sergiu, Cercetător Post-doctorand norma partiala
  • Ciocan Cristina, Cercetător Post-doctorand
  • Raduly Lajos, Cercetător Post-doctorand
  • Pirlog Radu, Cercetător Post-doctorand
  • Gherman Madalina, Student Doctorand
  • Muntean Maximilian Vlad, Cercetător Post-doctorand
  • 1 poziție vacantă, Cercetător Post-doctorand
  • Gheorghe-Milea Ana- Student doctorand 1
  • Cebotaru Iunia Patricia-Student doctorand 2
  • Isachesku Ekaterina-Student doctorand 3
  • Student doctorand 4- poziție vacantă
  • Sorițău Olga -Cercetător cu experiență
  • Cismaru Cosmin Andrei, Cercetător Post-doctorand
  • Zamfir Paula – Tehnician cercetare 1
  • Chiroi Paul – Tehnician cercetare 2
  • Iova Olga Maria – Tehnician cercetare 3
  • Muntean Ioan Mihai- Tehnician cercetare 4

 

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