Environmental insults are directly involved in cancer development. In particular, Ultraviolet (UV) radiation has been associated to the acquisition of different types of skin cancer and premature skin aging. UV radiation causes modifications in the genetic material of cells (DNA) that if not repaired properly will lead to a mutated DNA (mutated genes) which might trigger the development of cancer. Understanding the molecular basis of the UV-induced DNA damage response is important to elucidate the mechanisms of skin tumorigenesis.

In the study, published in Plos Genetics, the authors used a UV-induced skin cancer mouse model (Hepatocyte growth factor (HGF) transgenic mice), where one of the two Lkb1 gene alleles was deleted (haploinsufficiency), and consequently the amounts of LKB1 protein was half of the normal levels. A single dose of UVB radiation in Lkb1 haploinsufficient neonates  mice expressing HGF was enough to induced the quickly development of squamous cell carcinomas, and this, was associated to a deficient response in DNA damage repair.  Moreover, cells harboring the damaged DNA were resistant to cell death (apoptosis).  Thus, lack of LKB1 promotes a double effect: cells not only fail to repair the damage in their DNA, but they do not die, leading to the accumulation of mutated cells and the development of tumors. The research team has obtained similar results (to be published soon) in two additional mouse models of melanoma, a much more lethal type of UV-induced skin cancer.

One of the beauties of this model is that reflects the real scenario for cancer adquisition where initially just one of gene copies is damaged. It also reproduces a tumor related to chronic DNA damage using a single dose of UV radiation, underlining the importance of this protein’s function. 

LKB1 mutations have been found epithelial human cancers associated to environmental insults. “However, this is the first study that links LKB1 to the UV-induced DNA repair responses” explains Dr. Recio, “and provides an important insight into how cells can defend themselves from external genotoxic damage”.

Importantly, these results can be extrapolated to human cancer. In fact, when examining the expression of LKB1 protein in samples from patients with skin tumors, the authors found that roughly half of these samples showed low or no expression of LKB1.  Furthermore, absence of the protein was detected in all stages of the disease, particularly in UV-exposed skin areas, suggesting that the loss of expression of LKB1 is an early event and very likely contributes to UV-induced skin cancer development.

In the near future, we will be evaluating LKB1 as a prognostic risk factor for UV–induced skin cancer. They also are investigating the different factors that may alter LKB1 expression, with a particular emphasis in families with skin cancer predisposition or skin cancer history.

Environmental insults are directly involved in cancer development. In particular, Ultraviolet (UV) radiation has been associated to the acquisition of different types of skin cancer and premature skin aging. UV radiation causes modifications in the genetic material of cells (DNA) that if not repaired properly will lead to a mutated DNA (mutated genes) which might trigger the development of cancer. Understanding the molecular basis of the UV-induced DNA damage response is important to elucidate the mechanisms of skin tumorigenesis.

In the study, published in Plos Genetics, the authors used a UV-induced skin cancer mouse model (Hepatocyte growth factor (HGF) transgenic mice), where one of the two Lkb1 gene alleles was deleted (haploinsufficiency), and consequently the amounts of LKB1 protein was half of the normal levels. A single dose of UVB radiation in Lkb1 haploinsufficient neonates  mice expressing HGF was enough to induced the quickly development of squamous cell carcinomas, and this, was associated to a deficient response in DNA damage repair.  Moreover, cells harboring the damaged DNA were resistant to cell death (apoptosis).  Thus, lack of LKB1 promotes a double effect: cells not only fail to repair the damage in their DNA, but they do not die, leading to the accumulation of mutated cells and the development of tumors. The research team has obtained similar results (to be published soon) in two additional mouse models of melanoma, a much more lethal type of UV-induced skin cancer.

One of the beauties of this model is that reflects the real scenario for cancer adquisition where initially just one of gene copies is damaged. It also reproduces a tumor related to chronic DNA damage using a single dose of UV radiation, underlining the importance of this protein’s function. 

LKB1 mutations have been found epithelial human cancers associated to environmental insults. “However, this is the first study that links LKB1 to the UV-induced DNA repair responses” explains Dr. Recio, “and provides an important insight into how cells can defend themselves from external genotoxic damage”.

Importantly, these results can be extrapolated to human cancer. In fact, when examining the expression of LKB1 protein in samples from patients with skin tumors, the authors found that roughly half of these samples showed low or no expression of LKB1.  Furthermore, absence of the protein was detected in all stages of the disease, particularly in UV-exposed skin areas, suggesting that the loss of expression of LKB1 is an early event and very likely contributes to UV-induced skin cancer development.

In the near future, we will be evaluating LKB1 as a prognostic risk factor for UV–induced skin cancer. They also are investigating the different factors that may alter LKB1 expression, with a particular emphasis in families with skin cancer predisposition or skin cancer history.

LABORATORY OF NEURAL TUMORS

Principal Investigator (Name, MD/PhD)

Miguel F. Segura

Research Team

María José Pérez García, Postdoctoral Researcher & Project Manager

Khloud Abdo Elsharawy, Postdoctoral Researcher

Isabel de Rojas de Pablo, Postdoctoral Researcher

Adrià Molero Valenzuela, PhD Researcher

Marta Miera Maluenda, PhD Researcher

María Gallego Rodríguez, PhD Researcher

Gal·la Farreny Fernández, PhD Researcher

María Jesús Rodríguez Rodríguez, Laboratory Technician

Clinical Associated Researchers

Lucas Moreno, MD PhD. Head of the Childhood Cancer and Blood Disorders Group. Expertise in drug development and clinical trials.

Anna Llort, MD PhD. Pediatric Oncologist and institutional lead for pediatric brain tumors.

María Pérez-Torres, MD. Pediatric Oncologist.

Mariona Morell Daniel, MD. Pediatric Oncologist.

Marina Ortiz Palacios, MD. Pediatric Oncologist.

BACKGROUND

Pediatric cancer is a distinct disease entity with biological and clinical features that differ substantially from those of adult tumors, including etiology, tumor biology, treatment response, and prognosis. Over the past two decades, the implementation of multimodal treatment strategies—combining chemotherapy, radiotherapy, surgery, and targeted agents—has increased overall survival to around 80%. However, this progress has not benefited all patients equally. Children with certain tumor types continue to face poor outcomes, and many survivors experience severe long-term treatment-related toxicities. Developing more effective therapies for patients with high-risk disease, while reducing the short- and long-term side effects of current treatments, therefore remains an urgent clinical need.

Tumors of the nervous system are the most common solid malignant tumors in childhood and the leading cause of cancer-related death in children. Our laboratory is primarily focused on the development and implementation of epigenetic therapies, which target regulators of gene expression and, in turn, modulate multiple genes, pathways, and cellular processes without altering the DNA sequence itself. We study both neuroblastoma, a pediatric solid tumor of the peripheral nervous system, and pediatric brain tumors of the central nervous system, including ependymoma, medulloblastoma, and high-grade glioma, among others. Moreover, we are expanding our research toward more translational approaches, including projects linked to the initiation of clinical trials and the collection of liquid biopsy samples, in order to facilitate the transfer of our findings into clinical applications for pediatric cancer patients.

RESEARCH STRATEGY AND SCOPE

The research strategy of the Neural Tumors Lab is based on the following research lines:

Unveiling the epigenetic drivers of metastatic neuroblastoma

Approximately half of neuroblastoma patients present with metastatic disease at diagnosis, most frequently involving bone, bone marrow, lymph nodes, liver, and lungs, being metastatic relapse the leading cause of death of the disease. Increasing evidence indicates that epigenetic deregulation plays a critical role in neuroblastoma initiation, progression, and metastatic dissemination. We therefore hypothesize that systematic characterization of chromatin architecture alterations and therapeutic targeting of epigenetic vulnerabilities will advance our understanding of neuroblastoma metastasis and uncover novel therapeutic opportunities.

Uncovering epigenetic vulnerabilities of neuroblastoma

Epigenetics determines which genes are turned on or off without altering the DNA sequence, largely by controlling how DNA is packaged within the cell. A key player in this process is the BAF complex, a chromatin remodeler that neuroblastoma cells rely on to sustain proliferation and metastatic potential. Recent work in our laboratory has identified ARID1A/ARID1B as a promising vulnerability within the BAF complex, and this opened up two new research branches: (1) The development of small molecules to disrupt this dependency and inhibit tumor growth and (2) the use of PROTACs and specific inhibitors of SMARCA4 as a synthetic lethality strategy for ARID1A/B mutated patients.

Drug development: Moving forward to early-phase clinical trials

We are interested in testing the therapeutic potential of new synthetic or natural compounds which can represent clear advantages (i.e. increased effectiveness, reduced toxicities) compared to traditional chemotherapy. We have a clinically representative panel of pediatric solid tumor cell lines and preclinical mouse models to test the efficacy and safety of new drugs aimed to improve the efficacy and safety of currently available treatments. Currently, we are evaluating the therapeutic potential of ABTL0812 in pediatric tumors, both preclinically but also in a “first in child” Phase I clinical trial.

Translational precision oncology for pediatric neural tumors: Liquid biopsy

Liquid biopsy is a minimally invasive approach that enables the analysis of tumor-derived material in biofluids such as blood. In this project, we investigate circulating tumor DNA (ctDNA) as a biomarker for the molecular characterization and clinical monitoring of pediatric brain tumors and neuroblastoma. ctDNA analysis can provide clinically relevant information on tumor burden, tumor-associated genetic alterations, and disease dynamics over time. By allowing serial sampling, this strategy offers an attractive alternative to repeated invasive procedures. Our aim is to evaluate the potential of ctDNA to improve diagnosis, support treatment monitoring, and enable earlier detection of relapse. Ultimately, this work seeks to advance more precise, dynamic, and less invasive approaches for the management of pediatric malignancies.

Advancing in brain tumors research

Because pediatric brain tumors are the most common solid malignancies in children and still have limited treatment options resulting in poor overall survival, one of the laboratory’s main objectives is to develop new therapeutic strategies for these tumors. To this end, we are generating novel preclinical models of pediatric ependymoma to better recapitulate tumor biology and support translational research and evaluating the therapeutic potential of novel experimental compounds in vitro, including SWI/SNF inhibitors currently explored in our neuroblastoma research lines. Moreover, we aim to advance immunotherapy in pediatric brain tumors by identifying tumor-specific membrane proteins that can serve as targets for the development of more precise and effective therapeutic strategies.

ONGOING COMPETITIVE PROJECTS:

Ref. PI23/01144. Exploiting epigenetic vulnerabilities in metastatic neuroblastoma. Instituto de Salud Carlos III. PI: Miguel F. Segura.

Summary: Neuroblastoma is a cancer of the sympathetic nervous system, and is the most common solid tumor of childhood, representing ~15% of cancer deaths in children. While patients with localized disease have seen significant advances in their outcome, improvement in survival for patients with metastatic disease has been more limited. The liver, bone and bone marrow and lymph nodes are among the most common metastatic sites of neuroblastoma patients, which constitute a “reservoir” of tumor cells that persistently reside in patients following local and systemic cancer therapy. Their elimination continues to represent the most difficult challenges for neuroblastoma patients. Furthermore, the lack of clinically-representative models of neuroblastoma metastasis is hindering the development of therapies to target this subpopulation of cells. Our hypothesis is that metastatic cells can colonize distant organs thanks to the re-organization of their epigenetic landscape thanks to chromatin remodelers such as the SWI/SNF complex. We plan to validate our hypothesis through the following objectives: i) characterization of the epigenetic landscape of metastatic neuroblastoma; ii) characterize the transcriptomic map of SWI/SNF-regulated genes in neuroblastoma metastasis; iii) development of an epigenetic therapy based on the inhibition of chromatin remodeling complexes iv) development of more sensitive tools to diagnose and target neuroblastoma metastasis. To complete these tasks, we will use unique neuroblastoma metastasis mouse models combined with genomic analyses (ATACseq, RNAseq) and development of new compounds (SWI/SNF inhibitors) and diagnostic tools.

Ref. MONALISA_HE-MISS2023. MONALISA: A SIOPEN pragmatic clinical trial to MOnitor NeuroblastomA relapse with LIquid biopsy Sensitive Analysis. PI: Lucas Moreno.

Summary: High-risk neuroblastoma accounts for 15% of cancer related-deaths in children. Half of the >1500 patients yearly diagnosed with neuroblastoma in the EU have high-risk disease, which will relapse or progress in half these cases after first-line treatment. Relapsed neuroblastoma is aggressive and often therapy-resistant. Monitoring for disease relapse and therapy response is crucial for the survival chance of these patients. The current standard-of-care for monitoring are imaging technologies and bone marrow assessment, which are costly, invasive and a burden for children, who often require anesthesia. These drawbacks limit how often is monitored. More sensitive, less invasive and less toxic monitoring techniques are needed. The mutational spectrum often changes in recurring tumors, which may explain therapy resistance and provide additional druggable targets. Imaging, however, provides no information about molecular characteristics. Liquid biopsy tests are minimally invasive, allow frequent sampling and sensitively detect tumor molecular markers in tumor-derived DNA and messenger RNA circulating in peripheral blood. MONALISA aims to close existing gaps and establish liquid biopsies as standard-of-care to monitor relapsed/refractory neuroblastoma, as a blueprint for other pediatric cancers. Reliable, early assessment of molecular progression or relapse is the main aim of the pragmatic randomized clinical trial proposed in MONALISA. We develop a digital decision support tool to help oncologists use the new monitoring and apply patient-reported outcomes to integrate patient viewpoints and assess the effect of minimally invasive, liquid biopsy diagnostics on quality of life. We will establish whether events can be detected earlier using liquid biopsy monitoring, and whether better overall survival is enabled by earlier diagnosis and treatment interventions. This essential step towards personalized medicine will support reliable disease monitoring under treatment. “This action is part of the Cancer Mission cluster of projects on ‘‘Diagnostics and Treatment (diagnostics).

Ref. HRCI/JOINT_FUNDING_SCHEME/2022/SEGURA. Development of mRNA vaccines for children with high-risk neuroblastoma.

Summary: Neuroblastoma is one of the most aggressive childhood cancers contributing to 15% of cancer related childhood deaths. At diagnosis half of the patients have a metastatic tumour and recurrence is very common. Despite advances in available therapies, children with drug-resistant and relapsed neuroblastoma have a dismal outlook with 5-year survival rates of less than 20%, highlighting the need for new treatments. Immunotherapies, including anti-tumour vaccines, hold great promise to effectively target the tumour while generating fewer side effects and associated toxicities. This is achieved through the mechanism of action of vaccines that teach the immune system to identify and combat the tumour offering long term protection against recurrence. It is also forecasted that the success of COVID-19 messenger ribonucleic acid (mRNA) vaccines is set to boost the therapeutic oligonucleotide market to reach $4.5B by 2027 globally. This project aims to develop the first mRNA vaccine against high-risk neuroblastoma. Specifically, we will select two well characterised neuroblastoma cell surface antigens to be targeted in a mRNA vaccine. This vaccine will be delivered using RALA peptide, based on previous studies of the applicant team. The RALA/mRNA nanoparticles will be characterized for shape, surface charge, stability and immunogenicity. The transfection efficiency of the vaccine will be determined in vitro using DC 2.4 cells (murine dendritic cell line) and in THP-1 cells (human monocytic cell line). For the immunogenicity studies, C57 BL/6 mice will receive the vaccine and blood samples will be collected to analyse specific antibodies and cytokine secretion. The therapeutic and prophylactic potential of RALA/mRNA vaccine will be analysed using a metastatic model of neuroblastoma and a PDX humanized mouse model. Finally, we will evaluate the regulatory options to ensure an efficient therapy translation to the clinical setting. This will include market analysis as well as intellectual property and orphan drug designation applications.

Ref. 2024 PROD 00054. Targeting chromatin remodelers for cancer therapy.

Summary: Neuroblastoma (NB), a pediatric tumor, poses significant challenges due to its high mortality rates and resistance to conventional treatments. Many high-risk patients do not achieve a cure, and even survivors may suffer severe long-term side effects. Current therapies often fall short in cases of relapse or disease progression.

NB's ability to metastasize and treatment resistance involves genomic reprogramming facilitated by aberrant function of epigenetic regulators such as the mSWI/SNF chromatin remodeling complex. Lately, our group have made crucial discoveries, linking elevated expression of specific subunits (e.g., SMARCA4) with poor prognosis. Functionally, we found that by silencing only two subunits (ARID1A/B) the complex was destabilized, which inhibited tumor growth and metastasis. Inhibition of protein-protein interactions in the SWI/SNF complex innovates in the development of a first-in-class family of drugs. These compounds demonstrated anti-proliferative properties across a wide range of NB cell lines, with high selectivity and minimal impact on cells lacking ARID1A/B subunits. The main objective of this project is to lead optimization of our novel compounds, study their efficacy in clinically-representative NB models and establish an IP and a commercial strategy for the transition of this family of compounds to the market.

Compared to the standard NB treatment, our strategy offers a distinct mechanism of action by reversing epigenetic alterations without causing DNA damage in healthy tissues. This approach promises enhanced treatment efficacy and reduced long-term side effects. Moreover, our solution may improve the quality of life in pediatric cancer patients, reducing the physical and emotional burden on affected families as well as improving the outcomes for high-risk NB cases. This novel epigenetic treatment will also contribute to the growth of the epigenetics market, which is projected to exhibit substantial growth in the coming years.

Ref: NeuVac – a multiepitope mRNA vaccine for high-risk neuroblastoma

Summary: Neuroblastoma is one of the most aggressive childhood cancers contributing to 15% of cancer related childhood deaths. At diagnosis half of the patients have a metastatic tumour and recurrence is very common. Despite advances in available therapies, children with drug-resistant and relapsed neuroblastoma have a dismal outlook with 5-year survival rates of less than 20%, highlighting the need for new treatments. Immunotherapies, including anti-tumour vaccines, hold great promise to effectively target the tumour while generating fewer side effects and associated toxicities. This is achieved through the mechanism of action of vaccines that teach the immune system to identify and combat the tumour offering long-term protection against recurrence. This project aims to design and manufacture a multiepitope mRNA vaccine against high-risk neuroblastoma. Specifically, we will characterise and shortlist neuroblastoma cell surface antigens to be targeted in a mRNA vaccine. This vaccine will be delivered using RALA peptide, based on previous studies of the applicant team. We will characterise the RALA/mRNA formulation, stability and assessment of immune response. We will identify an appropriate tumour model in vivo and determine the therapeutic response to the vaccine in a metastatic model of neuroblastoma in a clinically relevant scenario. Should these critical research activities provide positive results, we are then ideally placed to move to the next stage of the development process (toxicology) and this provides the data package to enable further application to the orphan drug designation application and first children clinical trials.

Ref. BARRERA/BECA/SEGURA/2024. Unlocking the Power of Liquid Biopsy: Advancing Precision Medicine in Pediatric Brain Tumors.

Summary: Los tumores del sistema nervioso central son el tumor sólido pediátrico más frecuente representando hasta el 20-25% de todos los casos. A pesar de los múltiples avances en oncohematología pediátrica, muchos de estos tumores siguen teniendo un pronóstico desfavorable. Las herramientas actuales para su diagnóstico y monitorización presentan limitaciones, tanto las pruebas de imagen, que pueden llevar a interpretaciones confusas especialmente después de la cirugía; como la biopsia del tejido, que no es representativa de la heterogeneidad tumoral y puede interferir en el neurodesarrollo, causando daños a largo plazo. Por estas razones es necesario innovar y desarrollar nuevos métodos de detección que permitan un abordaje clínico menos invasivo de estos tumores. La hipótesis de este proyecto es que el análisis del ADN tumoral circulante (ctDNA) obtenido en muestras de sangre o líquido cefalorraquídeo de los pacientes con tumores cerebrales pediátricos (TCP) puede proporcionar información clínicamente relevante, posicionándose como una herramienta útil y mínimamente invasiva para el diagnóstico, monitorización y seguimiento de estos casos. Esta hipótesis se validará mediante los siguientes objetivos:

i. Obtención de muestras y recogida de casos retrospectivos. ii. Identificación de biomarcadores representativos en los distintos tipos de TCPs. iii. Prueba piloto para la validación y optimización de la técnica de detección.

Estas tareas se completarán mediante la recolección de muestras biológicas y el uso de tecnologías avanzadas como PCR digital y secuenciación de nueva generación (NGS) para detectar alteraciones moleculares recurrentes del ctDNA de las muestras. Este proyecto tiene el potencial de transformar el diagnóstico y seguimiento de los tumores cerebrales pediátricos, mejorando la supervivencia y calidad de vida de los niños afectados, al tiempo que ofrece esperanza y bienestar a sus familias.

Ref. 2025 LLAV 00002. AURA-BRAIN: AI-powered Utility for Real-time Assessment and decision-making in pediatric Brain tumors using liquid biopsy.

Summary: represent the most common solid group of cancers in children, accounting for approximately 25% of all pediatric tumors. Despite advances in therapeutic strategies, PBTs remain a leading cause of cancer-related mortality in children. A majority of patients experience sudden tumor progression, therapy resistance, and/or relapse, underscoring the urgent need for more effective follow-up and intervention protocols.

Current monitoring approaches, such as imaging, are unable to provide molecular insights into tumor evolution, frequently exhibiting altered mutational landscapes, which are critical for addressing therapy resistance and identifying new druggable targets. In this sense, liquid biopsy has emerged as a promising minimally invasive technique, capable of detecting circulating-tumor DNA (ctDNA) in peripheral blood and cerebrospinal fluid (CSF). This approach offers the potential for frequent sampling, sensitive detection of tumor biomarkers, and real-time insights into tumor dynamics. In this proposal, we submit a project that aims to translate current PBT research knowledge into clinical practice by developing liquid biopsy technologies that will enable better clinical management of patients at diagnosis, treatment and monitoring stages. The primary objective is to develop and validate a next generation sequencing (NGS)-based protocol using circulating tumor nucleic acids as biomarkers. This technology will be supported by an artificial intelligence (AI) decision support system that integrates both genomic and clinical parameters to evaluate clinical outcomes. Its goal is to provide more accurate and less invasive methods for molecular diagnosis, tumor burden monitoring, treatment response assessment, disease progression tracking, and early relapse detection, ultimately improving survival outcomes for children with brain tumors.

Ref: HORIZON-MISS-2025-02-CANCER-04-101289276. Phase I/II trial of an Oral ER-stress iNducer in relapsed/refractory neuroblastoma and paediatric solid tumours.

Summary: Childhood cancer is the leading cause of disease-related death in children, with high-risk neuroblastoma and other aggressive paediatric solid tumours representing a critical unmet need. Despite multimodal therapy, over half of children with relapsed or refractory disease do not achieve long-term survival, and those who do often suffer severe, lifelong toxicities. There is an urgent need for safer and more effective therapies.

Ibrilatazar (ABTL0812) is a first-in-class, orally administered anticancer agent that induces tumour cell death through endoplasmic reticulum stress and cytotoxic autophagy. Unlike conventional genotoxic chemotherapies, it spares DNA, offering a potentially safer profile highly relevant in children. In adult patients with advanced solid tumours, ibrilatazar has shown an excellent safety record and early efficacy in Phase I/II trials, with long-term stabilisation in heavily pre-treated patients, thereby supporting its evaluation in the paediatric population.

Preclinical studies in neuroblastoma models, including MYCN-amplified and chemoresistant cell lines, demonstrate that ibrilatazar reduces tumour growth, downregulates MYCN expression, and enhances standard chemotherapies and differentiating agents.

Building on this evidence, PHOENIX will conduct a first-in-child, multicentre Phase I/II clinical trial of ibrilatazar in relapsed/refractory neuroblastoma and other aggressive solid tumours. The trial will test combinations with irinotecan-temozolomide and selected immunotherapies (anti-GD2, anti-VEGF), aligned with our preclinical findings. In parallel, a comprehensive translational programme will integrate pharmacokinetic, pharmacodynamic, genomic, and immunological biomarker studies, aiming to enable patient stratification, early treatment monitoring, and improved trial inclusiveness by addressing social determinants of health. Thus, PHOENIX seeks to deliver a safer, more effective therapeutic alternative for children with high-risk cancers.

SELECTED PUBLICATIONS

1. Miera-Maluenda M, Pérez-Torres M, Mañas A, Rubio-San-Simón A, Butjosa-Espín M, Ruiz-Duran P, Seoane JA, Moreno L, Segura MF. Advances in the approaches used to repurpose drugs for neuroblastoma. Expert Opin Drug Discov. 2024 Nov;19(11):1309-1319. doi: 10.1080/17460441.2024.2402413. Epub 2024 Sep 11. PMID: 39258785.

2. Murphy C, Devis-Jauregui L, Struck R, Boloix A, Gallagher C, Gavin C, Cottone F, Fernandez AS, Madden S, Roma J, Segura MF*, Piskareva O*. In vivo cisplatin-resistant neuroblastoma metastatic model reveals tumour necrosis factor receptor superfamily member 4 (TNFRSF4) as an independent prognostic factor of survival in neuroblastoma. PLoS One. 2024 May 29;19(5):e0303643. doi: 10.1371/journal.pone.0303643. PMID: 38809883; PMCID: PMC11135766. (* corresponding authors).

3. Pérez-García MJ, Segura MF. Maintaining excellent outcomes: the impact of age cutoff reclassification on reduced therapy for neuroblastoma patients. Transl Pediatr. 2023 Nov 28;12(11):1926-1930. doi: 10.21037/tp-23-391. Epub 2023 Nov 23. Erratum in: Transl Pediatr. 2024 Aug 31;13(8):1514. doi: 10.21037/tp-2024-02. PMID: 38130585; PMCID: PMC10730960.

4. Jiménez C, Moreno L, Segura MF. Epigenetic therapies for neuroblastoma: immunogenicity awakens. Mol Oncol. 2023 May;17(5):718-721. doi: 10.1002/1878-0261.13404. Epub 2023 Mar 8. PMID: 36840349; PMCID: PMC10158771.

5.- Jiménez C, Antonelli R, Nadal-Ribelles M, Devis-Jauregui L, Latorre P, Solé C, Masanas M, Molero-Valenzuela A, Soriano A, Sánchez de Toledo J, Llobet-Navas D, Roma J, Posas F, de Nadal E, Gallego S, Moreno L, Segura MF. Structural disruption of BAF chromatin remodeller impairs neuroblastoma metastasis by reverting an invasiveness epigenomic program. Mol Cancer. 2022 Sep 3;21(1):175. doi: 10.1186/s12943-022-01643-4. PMID: 36057593; PMCID: PMC9440539.

6.- Segura MF, Soriano A, Roma J, Piskareva O, Jiménez C, Boloix A, Fletcher JI, Haber M, Gray JC, Cerdá-Alberich L, Martínez de Las Heras B, Cañete A, Gallego S, Moreno L. Methodological advances in the discovery of novel neuroblastoma therapeutics. Expert Opin Drug Discov. 2022 Feb;17(2):167-179. doi: 10.1080/17460441.2022.2002297. Epub 2021 Nov 22. PMID: 34807782.

7.- Boloix A, Feiner-Gracia N, Köber M, Repetto J, Pascarella R, Soriano A, Masanas M, Segovia N, Vargas-Nadal G, Merlo-Mas J, Danino D, Abutbul-Ionita I, Foradada L, Roma J, Córdoba A, Sala S, de Toledo JS, Gallego S, Veciana J, Albertazzi L, Segura MF*, Ventosa N*. Engineering pH-Sensitive Stable Nanovesicles for Delivery of MicroRNA Therapeutics. Small. 2022 Jan;18(3):e2101959. doi: 10.1002/smll.202101959. Epub 2021 Nov 16. PMID: 34786859. (*corresponding authors).

8.- Masanas M, Masiá N, Suárez-Cabrera L, Olivan M, Soriano A, Majem B, Devis-Jauregui L, Burgos-Panadero R, Jiménez C, Rodriguez-Sodupe P, Boloix A, Toledano I, Guillén G, Navarro A, Llobet-Navas D, Villanueva A, Sánchez de Toledo J, Roma J, Noguera R, Moreno L, Krauss R, Gallego S, Santamaria A*, Segura MF*. The oral KIF11 inhibitor 4SC-205 exhibits antitumor activity and potentiates standard and targeted therapies in primary and metastatic neuroblastoma models. Clin Transl Med. 2021 Oct;11(10):e533. doi: 10.1002/ctm2.533. PMID: 34709738; PMCID: PMC8516339. (*corresponding authors).

9.- París-Coderch L, Soriano A, Jiménez C, Erazo T, Muñoz-Guardiola P, Masanas M, Antonelli R, Boloix A, Alfón J, Pérez-Montoyo H, Yeste-Velasco M, Domènech C, Roma J, Sánchez de Toledo J, Moreno L, Lizcano JM, Gallego S, Segura MF. The antitumour drug ABTL0812 impairs neuroblastoma growth through endoplasmic reticulum stress-mediated autophagy and apoptosis. Cell Death Dis. 2020 Sep 17;11(9):773. doi: 10.1038/s41419-020-02986-w. PMID: 32943619; PMCID: PMC7498451.

10.- Antonelli R, Jiménez C, Riley M, Servidei T, Riccardi R, Soriano A, Roma J, Martínez-Saez E, Martini M, Ruggiero A, Moreno L, Sánchez de Toledo J, Gallego S, Bové J, Hooker JM, Segura MF. CN133, a Novel Brain-Penetrating Histone Deacetylase Inhibitor, Hampers Tumor Growth in Patient-Derived Pediatric Posterior Fossa Ependymoma Models. Cancers (Basel). 2020 Jul 16;12(7):1922. doi: 10.3390/cancers12071922. PMID: 32708733; PMCID: PMC7409080.

11.- Qadeer ZA, Valle-Garcia D, Hasson D, Sun Z, Cook A, Nguyen C, Soriano A, Ma A, Griffiths LM, Zeineldin M, Filipescu D, Jubierre L, Chowdhury A, Deevy O, Chen X, Finkelstein DB, Bahrami A, Stewart E, Federico S, Gallego S, Dekio F, Fowkes M, Meni D, Maris JM, Weiss WA, Roberts SS, Cheung NV, Jin J, Segura MF, Dyer MA, Bernstein E. ATRX In-Frame Fusion Neuroblastoma Is Sensitive to EZH2 Inhibition via Modulation of Neuronal Gene Signatures. Cancer Cell. 2019 Nov 11;36(5):512-527.e9. doi: 10.1016/j.ccell.2019.09.002. Epub 2019 Oct 17. PMID: 31631027; PMCID: PMC6851493.

12.- Soriano A, Masanas M, Boloix A, Masiá N, París-Coderch L, Piskareva O, Jiménez C, Henrich KO, Roma J, Westermann F, Stallings RL, Sábado C, de Toledo JS, Santamaria A, Gallego S, Segura MF. Functional high-throughput screening reveals miR-323a-5p and miR-342-5p as new tumor-suppressive microRNA for neuroblastoma. Cell Mol Life Sci. 2019 Jun;76(11):2231-2243. doi: 10.1007/s00018-019-03041-4. Epub 2019 Feb 15. PMID: 30770954; PMCID: PMC6502783.

13.- Jubierre L, Soriano A, Planells-Ferrer L, París-Coderch L, Tenbaum SP, Romero OA, Moubarak RS, Almazán-Moga A, Molist C, Roma J, Navarro S, Noguera R, Sánchez-Céspedes M, Comella JX, Palmer HG, Sánchez de Toledo J, Gallego S, Segura MF. BRG1/SMARCA4 is essential for neuroblastoma cell viability through modulation of cell death and survival pathways. Oncogene. 2016 Sep 29;35(39):5179-90. doi: 10.1038/onc.2016.50. Epub 2016 Mar 21. PMID: 26996667.

ACTIVE COLLABORATIONS WITH INDUSTRY: (Name, country)

Ability Pharma S.L. (Barcelona, Spain)

OMICA Biomed S.L.

PAST MEMBERS: (Name, actual position)

-Dr. Luz Jubierre Zapater (2013-2017). Senior Research Scientist at MSKCC, NY, USA.

-Dr. Laia París-Coderch (2014-2017). Currently working as Validation and GMP Junior Consultant at TDV SL.

-Dr. Aroa Soriano (2012-2020). Currently leading the Personalized Medicine Program at Vall Hebron Hospital.

-Dr. Marc Masanas (2015-2022). High-school Teacher.

-Dr. Roberta Antonelli (2016-2022). Research and Development Senior Scientist and Scientific Project Manager at the Institute of Microelectronics of Barcelona (IMB-CNM, CSIC), Barcelona.

-Dr. Carlos Jiménez Jiménez (2015-2022), Postdoctoral Researcher at the Human Technopole Center (Milan).

LABORATORY OF NEURAL TUMORS

Principal Investigator (Name, MD/PhD)

Miguel F. Segura

Research Team

María José Pérez García, Postdoctoral Researcher & Project Manager

Khloud Abdo Elsharawy, Postdoctoral Researcher

Isabel de Rojas de Pablo, Postdoctoral Researcher

Adrià Molero Valenzuela, PhD Researcher

Marta Miera Maluenda, PhD Researcher

María Gallego Rodríguez, PhD Researcher

Gal·la Farreny Fernández, PhD Researcher

María Jesús Rodríguez Rodríguez, Laboratory Technician

Clinical Associated Researchers

Lucas Moreno, MD PhD. Head of the Childhood Cancer and Blood Disorders Group. Expertise in drug development and clinical trials.

Anna Llort, MD PhD. Pediatric Oncologist and institutional lead for pediatric brain tumors.

María Pérez-Torres, MD. Pediatric Oncologist.

Mariona Morell Daniel, MD. Pediatric Oncologist.

Marina Ortiz Palacios, MD. Pediatric Oncologist.

BACKGROUND

Pediatric cancer is a distinct disease entity with biological and clinical features that differ substantially from those of adult tumors, including etiology, tumor biology, treatment response, and prognosis. Over the past two decades, the implementation of multimodal treatment strategies—combining chemotherapy, radiotherapy, surgery, and targeted agents—has increased overall survival to around 80%. However, this progress has not benefited all patients equally. Children with certain tumor types continue to face poor outcomes, and many survivors experience severe long-term treatment-related toxicities. Developing more effective therapies for patients with high-risk disease, while reducing the short- and long-term side effects of current treatments, therefore remains an urgent clinical need.

Tumors of the nervous system are the most common solid malignant tumors in childhood and the leading cause of cancer-related death in children. Our laboratory is primarily focused on the development and implementation of epigenetic therapies, which target regulators of gene expression and, in turn, modulate multiple genes, pathways, and cellular processes without altering the DNA sequence itself. We study both neuroblastoma, a pediatric solid tumor of the peripheral nervous system, and pediatric brain tumors of the central nervous system, including ependymoma, medulloblastoma, and high-grade glioma, among others. Moreover, we are expanding our research toward more translational approaches, including projects linked to the initiation of clinical trials and the collection of liquid biopsy samples, in order to facilitate the transfer of our findings into clinical applications for pediatric cancer patients.

RESEARCH STRATEGY AND SCOPE

The research strategy of the Neural Tumors Lab is based on the following research lines:

Unveiling the epigenetic drivers of metastatic neuroblastoma

Approximately half of neuroblastoma patients present with metastatic disease at diagnosis, most frequently involving bone, bone marrow, lymph nodes, liver, and lungs, being metastatic relapse the leading cause of death of the disease. Increasing evidence indicates that epigenetic deregulation plays a critical role in neuroblastoma initiation, progression, and metastatic dissemination. We therefore hypothesize that systematic characterization of chromatin architecture alterations and therapeutic targeting of epigenetic vulnerabilities will advance our understanding of neuroblastoma metastasis and uncover novel therapeutic opportunities.

Uncovering epigenetic vulnerabilities of neuroblastoma

Epigenetics determines which genes are turned on or off without altering the DNA sequence, largely by controlling how DNA is packaged within the cell. A key player in this process is the BAF complex, a chromatin remodeler that neuroblastoma cells rely on to sustain proliferation and metastatic potential. Recent work in our laboratory has identified ARID1A/ARID1B as a promising vulnerability within the BAF complex, and this opened up two new research branches: (1) The development of small molecules to disrupt this dependency and inhibit tumor growth and (2) the use of PROTACs and specific inhibitors of SMARCA4 as a synthetic lethality strategy for ARID1A/B mutated patients.

Drug development: Moving forward to early-phase clinical trials

We are interested in testing the therapeutic potential of new synthetic or natural compounds which can represent clear advantages (i.e. increased effectiveness, reduced toxicities) compared to traditional chemotherapy. We have a clinically representative panel of pediatric solid tumor cell lines and preclinical mouse models to test the efficacy and safety of new drugs aimed to improve the efficacy and safety of currently available treatments. Currently, we are evaluating the therapeutic potential of ABTL0812 in pediatric tumors, both preclinically but also in a “first in child” Phase I clinical trial.

Translational precision oncology for pediatric neural tumors: Liquid biopsy

Liquid biopsy is a minimally invasive approach that enables the analysis of tumor-derived material in biofluids such as blood. In this project, we investigate circulating tumor DNA (ctDNA) as a biomarker for the molecular characterization and clinical monitoring of pediatric brain tumors and neuroblastoma. ctDNA analysis can provide clinically relevant information on tumor burden, tumor-associated genetic alterations, and disease dynamics over time. By allowing serial sampling, this strategy offers an attractive alternative to repeated invasive procedures. Our aim is to evaluate the potential of ctDNA to improve diagnosis, support treatment monitoring, and enable earlier detection of relapse. Ultimately, this work seeks to advance more precise, dynamic, and less invasive approaches for the management of pediatric malignancies.

Advancing in brain tumors research

Because pediatric brain tumors are the most common solid malignancies in children and still have limited treatment options resulting in poor overall survival, one of the laboratory’s main objectives is to develop new therapeutic strategies for these tumors. To this end, we are generating novel preclinical models of pediatric ependymoma to better recapitulate tumor biology and support translational research and evaluating the therapeutic potential of novel experimental compounds in vitro, including SWI/SNF inhibitors currently explored in our neuroblastoma research lines. Moreover, we aim to advance immunotherapy in pediatric brain tumors by identifying tumor-specific membrane proteins that can serve as targets for the development of more precise and effective therapeutic strategies.

ONGOING COMPETITIVE PROJECTS:

Ref. PI23/01144. Exploiting epigenetic vulnerabilities in metastatic neuroblastoma. Instituto de Salud Carlos III. PI: Miguel F. Segura.

Summary: Neuroblastoma is a cancer of the sympathetic nervous system, and is the most common solid tumor of childhood, representing ~15% of cancer deaths in children. While patients with localized disease have seen significant advances in their outcome, improvement in survival for patients with metastatic disease has been more limited. The liver, bone and bone marrow and lymph nodes are among the most common metastatic sites of neuroblastoma patients, which constitute a “reservoir” of tumor cells that persistently reside in patients following local and systemic cancer therapy. Their elimination continues to represent the most difficult challenges for neuroblastoma patients. Furthermore, the lack of clinically-representative models of neuroblastoma metastasis is hindering the development of therapies to target this subpopulation of cells. Our hypothesis is that metastatic cells can colonize distant organs thanks to the re-organization of their epigenetic landscape thanks to chromatin remodelers such as the SWI/SNF complex. We plan to validate our hypothesis through the following objectives: i) characterization of the epigenetic landscape of metastatic neuroblastoma; ii) characterize the transcriptomic map of SWI/SNF-regulated genes in neuroblastoma metastasis; iii) development of an epigenetic therapy based on the inhibition of chromatin remodeling complexes iv) development of more sensitive tools to diagnose and target neuroblastoma metastasis. To complete these tasks, we will use unique neuroblastoma metastasis mouse models combined with genomic analyses (ATACseq, RNAseq) and development of new compounds (SWI/SNF inhibitors) and diagnostic tools.

Ref. MONALISA_HE-MISS2023. MONALISA: A SIOPEN pragmatic clinical trial to MOnitor NeuroblastomA relapse with LIquid biopsy Sensitive Analysis. PI: Lucas Moreno.

Summary: High-risk neuroblastoma accounts for 15% of cancer related-deaths in children. Half of the >1500 patients yearly diagnosed with neuroblastoma in the EU have high-risk disease, which will relapse or progress in half these cases after first-line treatment. Relapsed neuroblastoma is aggressive and often therapy-resistant. Monitoring for disease relapse and therapy response is crucial for the survival chance of these patients. The current standard-of-care for monitoring are imaging technologies and bone marrow assessment, which are costly, invasive and a burden for children, who often require anesthesia. These drawbacks limit how often is monitored. More sensitive, less invasive and less toxic monitoring techniques are needed. The mutational spectrum often changes in recurring tumors, which may explain therapy resistance and provide additional druggable targets. Imaging, however, provides no information about molecular characteristics. Liquid biopsy tests are minimally invasive, allow frequent sampling and sensitively detect tumor molecular markers in tumor-derived DNA and messenger RNA circulating in peripheral blood. MONALISA aims to close existing gaps and establish liquid biopsies as standard-of-care to monitor relapsed/refractory neuroblastoma, as a blueprint for other pediatric cancers. Reliable, early assessment of molecular progression or relapse is the main aim of the pragmatic randomized clinical trial proposed in MONALISA. We develop a digital decision support tool to help oncologists use the new monitoring and apply patient-reported outcomes to integrate patient viewpoints and assess the effect of minimally invasive, liquid biopsy diagnostics on quality of life. We will establish whether events can be detected earlier using liquid biopsy monitoring, and whether better overall survival is enabled by earlier diagnosis and treatment interventions. This essential step towards personalized medicine will support reliable disease monitoring under treatment. “This action is part of the Cancer Mission cluster of projects on ‘‘Diagnostics and Treatment (diagnostics).

Ref. HRCI/JOINT_FUNDING_SCHEME/2022/SEGURA. Development of mRNA vaccines for children with high-risk neuroblastoma.

Summary: Neuroblastoma is one of the most aggressive childhood cancers contributing to 15% of cancer related childhood deaths. At diagnosis half of the patients have a metastatic tumour and recurrence is very common. Despite advances in available therapies, children with drug-resistant and relapsed neuroblastoma have a dismal outlook with 5-year survival rates of less than 20%, highlighting the need for new treatments. Immunotherapies, including anti-tumour vaccines, hold great promise to effectively target the tumour while generating fewer side effects and associated toxicities. This is achieved through the mechanism of action of vaccines that teach the immune system to identify and combat the tumour offering long term protection against recurrence. It is also forecasted that the success of COVID-19 messenger ribonucleic acid (mRNA) vaccines is set to boost the therapeutic oligonucleotide market to reach $4.5B by 2027 globally. This project aims to develop the first mRNA vaccine against high-risk neuroblastoma. Specifically, we will select two well characterised neuroblastoma cell surface antigens to be targeted in a mRNA vaccine. This vaccine will be delivered using RALA peptide, based on previous studies of the applicant team. The RALA/mRNA nanoparticles will be characterized for shape, surface charge, stability and immunogenicity. The transfection efficiency of the vaccine will be determined in vitro using DC 2.4 cells (murine dendritic cell line) and in THP-1 cells (human monocytic cell line). For the immunogenicity studies, C57 BL/6 mice will receive the vaccine and blood samples will be collected to analyse specific antibodies and cytokine secretion. The therapeutic and prophylactic potential of RALA/mRNA vaccine will be analysed using a metastatic model of neuroblastoma and a PDX humanized mouse model. Finally, we will evaluate the regulatory options to ensure an efficient therapy translation to the clinical setting. This will include market analysis as well as intellectual property and orphan drug designation applications.

Ref. 2024 PROD 00054. Targeting chromatin remodelers for cancer therapy.

Summary: Neuroblastoma (NB), a pediatric tumor, poses significant challenges due to its high mortality rates and resistance to conventional treatments. Many high-risk patients do not achieve a cure, and even survivors may suffer severe long-term side effects. Current therapies often fall short in cases of relapse or disease progression.

NB's ability to metastasize and treatment resistance involves genomic reprogramming facilitated by aberrant function of epigenetic regulators such as the mSWI/SNF chromatin remodeling complex. Lately, our group have made crucial discoveries, linking elevated expression of specific subunits (e.g., SMARCA4) with poor prognosis. Functionally, we found that by silencing only two subunits (ARID1A/B) the complex was destabilized, which inhibited tumor growth and metastasis. Inhibition of protein-protein interactions in the SWI/SNF complex innovates in the development of a first-in-class family of drugs. These compounds demonstrated anti-proliferative properties across a wide range of NB cell lines, with high selectivity and minimal impact on cells lacking ARID1A/B subunits. The main objective of this project is to lead optimization of our novel compounds, study their efficacy in clinically-representative NB models and establish an IP and a commercial strategy for the transition of this family of compounds to the market.

Compared to the standard NB treatment, our strategy offers a distinct mechanism of action by reversing epigenetic alterations without causing DNA damage in healthy tissues. This approach promises enhanced treatment efficacy and reduced long-term side effects. Moreover, our solution may improve the quality of life in pediatric cancer patients, reducing the physical and emotional burden on affected families as well as improving the outcomes for high-risk NB cases. This novel epigenetic treatment will also contribute to the growth of the epigenetics market, which is projected to exhibit substantial growth in the coming years.

Ref: NeuVac – a multiepitope mRNA vaccine for high-risk neuroblastoma

Summary: Neuroblastoma is one of the most aggressive childhood cancers contributing to 15% of cancer related childhood deaths. At diagnosis half of the patients have a metastatic tumour and recurrence is very common. Despite advances in available therapies, children with drug-resistant and relapsed neuroblastoma have a dismal outlook with 5-year survival rates of less than 20%, highlighting the need for new treatments. Immunotherapies, including anti-tumour vaccines, hold great promise to effectively target the tumour while generating fewer side effects and associated toxicities. This is achieved through the mechanism of action of vaccines that teach the immune system to identify and combat the tumour offering long-term protection against recurrence. This project aims to design and manufacture a multiepitope mRNA vaccine against high-risk neuroblastoma. Specifically, we will characterise and shortlist neuroblastoma cell surface antigens to be targeted in a mRNA vaccine. This vaccine will be delivered using RALA peptide, based on previous studies of the applicant team. We will characterise the RALA/mRNA formulation, stability and assessment of immune response. We will identify an appropriate tumour model in vivo and determine the therapeutic response to the vaccine in a metastatic model of neuroblastoma in a clinically relevant scenario. Should these critical research activities provide positive results, we are then ideally placed to move to the next stage of the development process (toxicology) and this provides the data package to enable further application to the orphan drug designation application and first children clinical trials.

Ref. BARRERA/BECA/SEGURA/2024. Unlocking the Power of Liquid Biopsy: Advancing Precision Medicine in Pediatric Brain Tumors.

Summary: Los tumores del sistema nervioso central son el tumor sólido pediátrico más frecuente representando hasta el 20-25% de todos los casos. A pesar de los múltiples avances en oncohematología pediátrica, muchos de estos tumores siguen teniendo un pronóstico desfavorable. Las herramientas actuales para su diagnóstico y monitorización presentan limitaciones, tanto las pruebas de imagen, que pueden llevar a interpretaciones confusas especialmente después de la cirugía; como la biopsia del tejido, que no es representativa de la heterogeneidad tumoral y puede interferir en el neurodesarrollo, causando daños a largo plazo. Por estas razones es necesario innovar y desarrollar nuevos métodos de detección que permitan un abordaje clínico menos invasivo de estos tumores. La hipótesis de este proyecto es que el análisis del ADN tumoral circulante (ctDNA) obtenido en muestras de sangre o líquido cefalorraquídeo de los pacientes con tumores cerebrales pediátricos (TCP) puede proporcionar información clínicamente relevante, posicionándose como una herramienta útil y mínimamente invasiva para el diagnóstico, monitorización y seguimiento de estos casos. Esta hipótesis se validará mediante los siguientes objetivos:

i. Obtención de muestras y recogida de casos retrospectivos. ii. Identificación de biomarcadores representativos en los distintos tipos de TCPs. iii. Prueba piloto para la validación y optimización de la técnica de detección.

Estas tareas se completarán mediante la recolección de muestras biológicas y el uso de tecnologías avanzadas como PCR digital y secuenciación de nueva generación (NGS) para detectar alteraciones moleculares recurrentes del ctDNA de las muestras. Este proyecto tiene el potencial de transformar el diagnóstico y seguimiento de los tumores cerebrales pediátricos, mejorando la supervivencia y calidad de vida de los niños afectados, al tiempo que ofrece esperanza y bienestar a sus familias.

Ref. 2025 LLAV 00002. AURA-BRAIN: AI-powered Utility for Real-time Assessment and decision-making in pediatric Brain tumors using liquid biopsy.

Summary: represent the most common solid group of cancers in children, accounting for approximately 25% of all pediatric tumors. Despite advances in therapeutic strategies, PBTs remain a leading cause of cancer-related mortality in children. A majority of patients experience sudden tumor progression, therapy resistance, and/or relapse, underscoring the urgent need for more effective follow-up and intervention protocols.

Current monitoring approaches, such as imaging, are unable to provide molecular insights into tumor evolution, frequently exhibiting altered mutational landscapes, which are critical for addressing therapy resistance and identifying new druggable targets. In this sense, liquid biopsy has emerged as a promising minimally invasive technique, capable of detecting circulating-tumor DNA (ctDNA) in peripheral blood and cerebrospinal fluid (CSF). This approach offers the potential for frequent sampling, sensitive detection of tumor biomarkers, and real-time insights into tumor dynamics. In this proposal, we submit a project that aims to translate current PBT research knowledge into clinical practice by developing liquid biopsy technologies that will enable better clinical management of patients at diagnosis, treatment and monitoring stages. The primary objective is to develop and validate a next generation sequencing (NGS)-based protocol using circulating tumor nucleic acids as biomarkers. This technology will be supported by an artificial intelligence (AI) decision support system that integrates both genomic and clinical parameters to evaluate clinical outcomes. Its goal is to provide more accurate and less invasive methods for molecular diagnosis, tumor burden monitoring, treatment response assessment, disease progression tracking, and early relapse detection, ultimately improving survival outcomes for children with brain tumors.

Ref: HORIZON-MISS-2025-02-CANCER-04-101289276. Phase I/II trial of an Oral ER-stress iNducer in relapsed/refractory neuroblastoma and paediatric solid tumours.

Summary: Childhood cancer is the leading cause of disease-related death in children, with high-risk neuroblastoma and other aggressive paediatric solid tumours representing a critical unmet need. Despite multimodal therapy, over half of children with relapsed or refractory disease do not achieve long-term survival, and those who do often suffer severe, lifelong toxicities. There is an urgent need for safer and more effective therapies.

Ibrilatazar (ABTL0812) is a first-in-class, orally administered anticancer agent that induces tumour cell death through endoplasmic reticulum stress and cytotoxic autophagy. Unlike conventional genotoxic chemotherapies, it spares DNA, offering a potentially safer profile highly relevant in children. In adult patients with advanced solid tumours, ibrilatazar has shown an excellent safety record and early efficacy in Phase I/II trials, with long-term stabilisation in heavily pre-treated patients, thereby supporting its evaluation in the paediatric population.

Preclinical studies in neuroblastoma models, including MYCN-amplified and chemoresistant cell lines, demonstrate that ibrilatazar reduces tumour growth, downregulates MYCN expression, and enhances standard chemotherapies and differentiating agents.

Building on this evidence, PHOENIX will conduct a first-in-child, multicentre Phase I/II clinical trial of ibrilatazar in relapsed/refractory neuroblastoma and other aggressive solid tumours. The trial will test combinations with irinotecan-temozolomide and selected immunotherapies (anti-GD2, anti-VEGF), aligned with our preclinical findings. In parallel, a comprehensive translational programme will integrate pharmacokinetic, pharmacodynamic, genomic, and immunological biomarker studies, aiming to enable patient stratification, early treatment monitoring, and improved trial inclusiveness by addressing social determinants of health. Thus, PHOENIX seeks to deliver a safer, more effective therapeutic alternative for children with high-risk cancers.

SELECTED PUBLICATIONS

1. Miera-Maluenda M, Pérez-Torres M, Mañas A, Rubio-San-Simón A, Butjosa-Espín M, Ruiz-Duran P, Seoane JA, Moreno L, Segura MF. Advances in the approaches used to repurpose drugs for neuroblastoma. Expert Opin Drug Discov. 2024 Nov;19(11):1309-1319. doi: 10.1080/17460441.2024.2402413. Epub 2024 Sep 11. PMID: 39258785.

2. Murphy C, Devis-Jauregui L, Struck R, Boloix A, Gallagher C, Gavin C, Cottone F, Fernandez AS, Madden S, Roma J, Segura MF*, Piskareva O*. In vivo cisplatin-resistant neuroblastoma metastatic model reveals tumour necrosis factor receptor superfamily member 4 (TNFRSF4) as an independent prognostic factor of survival in neuroblastoma. PLoS One. 2024 May 29;19(5):e0303643. doi: 10.1371/journal.pone.0303643. PMID: 38809883; PMCID: PMC11135766. (* corresponding authors).

3. Pérez-García MJ, Segura MF. Maintaining excellent outcomes: the impact of age cutoff reclassification on reduced therapy for neuroblastoma patients. Transl Pediatr. 2023 Nov 28;12(11):1926-1930. doi: 10.21037/tp-23-391. Epub 2023 Nov 23. Erratum in: Transl Pediatr. 2024 Aug 31;13(8):1514. doi: 10.21037/tp-2024-02. PMID: 38130585; PMCID: PMC10730960.

4. Jiménez C, Moreno L, Segura MF. Epigenetic therapies for neuroblastoma: immunogenicity awakens. Mol Oncol. 2023 May;17(5):718-721. doi: 10.1002/1878-0261.13404. Epub 2023 Mar 8. PMID: 36840349; PMCID: PMC10158771.

5.- Jiménez C, Antonelli R, Nadal-Ribelles M, Devis-Jauregui L, Latorre P, Solé C, Masanas M, Molero-Valenzuela A, Soriano A, Sánchez de Toledo J, Llobet-Navas D, Roma J, Posas F, de Nadal E, Gallego S, Moreno L, Segura MF. Structural disruption of BAF chromatin remodeller impairs neuroblastoma metastasis by reverting an invasiveness epigenomic program. Mol Cancer. 2022 Sep 3;21(1):175. doi: 10.1186/s12943-022-01643-4. PMID: 36057593; PMCID: PMC9440539.

6.- Segura MF, Soriano A, Roma J, Piskareva O, Jiménez C, Boloix A, Fletcher JI, Haber M, Gray JC, Cerdá-Alberich L, Martínez de Las Heras B, Cañete A, Gallego S, Moreno L. Methodological advances in the discovery of novel neuroblastoma therapeutics. Expert Opin Drug Discov. 2022 Feb;17(2):167-179. doi: 10.1080/17460441.2022.2002297. Epub 2021 Nov 22. PMID: 34807782.

7.- Boloix A, Feiner-Gracia N, Köber M, Repetto J, Pascarella R, Soriano A, Masanas M, Segovia N, Vargas-Nadal G, Merlo-Mas J, Danino D, Abutbul-Ionita I, Foradada L, Roma J, Córdoba A, Sala S, de Toledo JS, Gallego S, Veciana J, Albertazzi L, Segura MF*, Ventosa N*. Engineering pH-Sensitive Stable Nanovesicles for Delivery of MicroRNA Therapeutics. Small. 2022 Jan;18(3):e2101959. doi: 10.1002/smll.202101959. Epub 2021 Nov 16. PMID: 34786859. (*corresponding authors).

8.- Masanas M, Masiá N, Suárez-Cabrera L, Olivan M, Soriano A, Majem B, Devis-Jauregui L, Burgos-Panadero R, Jiménez C, Rodriguez-Sodupe P, Boloix A, Toledano I, Guillén G, Navarro A, Llobet-Navas D, Villanueva A, Sánchez de Toledo J, Roma J, Noguera R, Moreno L, Krauss R, Gallego S, Santamaria A*, Segura MF*. The oral KIF11 inhibitor 4SC-205 exhibits antitumor activity and potentiates standard and targeted therapies in primary and metastatic neuroblastoma models. Clin Transl Med. 2021 Oct;11(10):e533. doi: 10.1002/ctm2.533. PMID: 34709738; PMCID: PMC8516339. (*corresponding authors).

9.- París-Coderch L, Soriano A, Jiménez C, Erazo T, Muñoz-Guardiola P, Masanas M, Antonelli R, Boloix A, Alfón J, Pérez-Montoyo H, Yeste-Velasco M, Domènech C, Roma J, Sánchez de Toledo J, Moreno L, Lizcano JM, Gallego S, Segura MF. The antitumour drug ABTL0812 impairs neuroblastoma growth through endoplasmic reticulum stress-mediated autophagy and apoptosis. Cell Death Dis. 2020 Sep 17;11(9):773. doi: 10.1038/s41419-020-02986-w. PMID: 32943619; PMCID: PMC7498451.

10.- Antonelli R, Jiménez C, Riley M, Servidei T, Riccardi R, Soriano A, Roma J, Martínez-Saez E, Martini M, Ruggiero A, Moreno L, Sánchez de Toledo J, Gallego S, Bové J, Hooker JM, Segura MF. CN133, a Novel Brain-Penetrating Histone Deacetylase Inhibitor, Hampers Tumor Growth in Patient-Derived Pediatric Posterior Fossa Ependymoma Models. Cancers (Basel). 2020 Jul 16;12(7):1922. doi: 10.3390/cancers12071922. PMID: 32708733; PMCID: PMC7409080.

11.- Qadeer ZA, Valle-Garcia D, Hasson D, Sun Z, Cook A, Nguyen C, Soriano A, Ma A, Griffiths LM, Zeineldin M, Filipescu D, Jubierre L, Chowdhury A, Deevy O, Chen X, Finkelstein DB, Bahrami A, Stewart E, Federico S, Gallego S, Dekio F, Fowkes M, Meni D, Maris JM, Weiss WA, Roberts SS, Cheung NV, Jin J, Segura MF, Dyer MA, Bernstein E. ATRX In-Frame Fusion Neuroblastoma Is Sensitive to EZH2 Inhibition via Modulation of Neuronal Gene Signatures. Cancer Cell. 2019 Nov 11;36(5):512-527.e9. doi: 10.1016/j.ccell.2019.09.002. Epub 2019 Oct 17. PMID: 31631027; PMCID: PMC6851493.

12.- Soriano A, Masanas M, Boloix A, Masiá N, París-Coderch L, Piskareva O, Jiménez C, Henrich KO, Roma J, Westermann F, Stallings RL, Sábado C, de Toledo JS, Santamaria A, Gallego S, Segura MF. Functional high-throughput screening reveals miR-323a-5p and miR-342-5p as new tumor-suppressive microRNA for neuroblastoma. Cell Mol Life Sci. 2019 Jun;76(11):2231-2243. doi: 10.1007/s00018-019-03041-4. Epub 2019 Feb 15. PMID: 30770954; PMCID: PMC6502783.

13.- Jubierre L, Soriano A, Planells-Ferrer L, París-Coderch L, Tenbaum SP, Romero OA, Moubarak RS, Almazán-Moga A, Molist C, Roma J, Navarro S, Noguera R, Sánchez-Céspedes M, Comella JX, Palmer HG, Sánchez de Toledo J, Gallego S, Segura MF. BRG1/SMARCA4 is essential for neuroblastoma cell viability through modulation of cell death and survival pathways. Oncogene. 2016 Sep 29;35(39):5179-90. doi: 10.1038/onc.2016.50. Epub 2016 Mar 21. PMID: 26996667.

ACTIVE COLLABORATIONS WITH INDUSTRY: (Name, country)

Ability Pharma S.L. (Barcelona, Spain)

OMICA Biomed S.L.

PAST MEMBERS: (Name, actual position)

-Dr. Luz Jubierre Zapater (2013-2017). Senior Research Scientist at MSKCC, NY, USA.

-Dr. Laia París-Coderch (2014-2017). Currently working as Validation and GMP Junior Consultant at TDV SL.

-Dr. Aroa Soriano (2012-2020). Currently leading the Personalized Medicine Program at Vall Hebron Hospital.

-Dr. Marc Masanas (2015-2022). High-school Teacher.

-Dr. Roberta Antonelli (2016-2022). Research and Development Senior Scientist and Scientific Project Manager at the Institute of Microelectronics of Barcelona (IMB-CNM, CSIC), Barcelona.

-Dr. Carlos Jiménez Jiménez (2015-2022), Postdoctoral Researcher at the Human Technopole Center (Milan).