Lorem Sit Amet Dolor
Researcher: Lorem Sit Amet

123 abc lane, Townsville, ZZ 00000, USA
Funding Progress: $§ / $§§§§§

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Exploiting the use of exosomal miRNAs from DIPG/DMG liquid biopsy as biomarkers of therapeutic response
Translational
DIPG, Childhood (Brain Cancer)
Lay Summary

An unmet need in DIPG/DMG, is  the lack of reliable biomarkers. Those are urgently needed as alternative/complementary approach to neuroradiological imaging, not only for diagnosis but for disease monitoring. In this context, liquid biopsy could represent a valuable, non-invasive source for biomarkers in DIPG/DMG.

From previous DIPG Collaborative funding, we have evidence that tumour exosomes, which are cellular microvescicles containing RNA, DNA, and proteins, are secreted from DIPG cells and can be found in patient plasma samples, carrying “biomolecules”  used to mediate the communication between tumor cells and/or tumor cells and the microenvironment.  

Based on this, we hypothesize that micro-RNA (small RNA molecules) carryed in the exosomes isolated from the liquid biopsy of DIPG/DMG patients, could be used as valid tumor biomarkers, providing information on therapeutic response, in real time through blood or CSF sample collections. To prove our hypothesis we will carry out:

- a prospective  study focused on the blood patient plasma samples which will take advantage of an ongoing clinical trial in our institutions.

- a retrospective  study based on the analysis on CSF derived exo-miRNAs

We expect our study will demonstrate the value of exosomal miRNAs from DIPG/DMG liquid biopsy as diagnostic/prognostic biomarkers and their potential application to clinical practice.

Executive Summary

Diffuse Intrinsic Pontine Glioma (DIPG) and Diffuse Midline Glioma H3-K27M mutant (commonly referred to as DMG) are the most aggressive tumors of the central nervous system for which there is no effective treatment, but also no reliable biomarkers.

The lack of biomarkers for DIPG/DMG represents an urgent unmet clinical need.

Despite the fact that biopsy procedures are more commonly practised, the centres where this procedure is offered are still a minority, and even when it takes place, it involves risks of morbidity or mortality which are not usually justifiable with better therapeutic options. The standard for DIPG diagnosis, but also to monitor disease progression, is still magnetic resonance imaging (MRI). Unfortunately this method cannot really distinguish between molecular subgroups of DIPG/DMG and also present with the difficulties to precisely differentiate tumour progression from pseudo-progression. The advent of liquid biopsy has open to the possibility to apply in the clinical practice non-invasive biomarkers. Depending on the cancer type, circulating-free tumour DNA (ct-DNA) and circulating tumour cells (CTC) have been reported to represent valuable analytes for detecting human malignancies at different stages of the disease by tracking driver mutations found in the primary tumor and/or the number of circulating cells. For brain tumors though, ctDNA and CTC have been used with variable success depending on the type of fluid, the volumes and the sequencing methods.

An alternative and attractive liquid biopsy analyte is represented by the exosomes, cellular microvescicles transporting different biomolecules, including DNA, RNA, miRNAs and proteins.

In previous funded project by DIPG Collaborative, we have initiated an innovative line of research studying the exosomes in DIPG and pHGG, given their known function in mediating cell-cell and cell-microenvironment cross-talk. Our data suggest that exosomes and in particular exosomal miRNAs (exo-miRNAs) can be used for pHGG/DIPG subgrouping based on the association of specific miRNAs signatures to specific driver mutations and/or tumor location.  MicroRNAs in general, including exo-miRNAs, are short, stable, single-stranded, noncoding RNAs that have been shown to control gene expression and being implicated in several tumour related properties like stemness and invasion and are considered attractive biomarkers for cancer. Interestingly, it has been shown that exo-miRNAs are distinct and superior when compared to “free-circulating” miRNAs, in preoperative diagnosis of glioblastoma.

Based on this we believe that exo-miRNAs have great potential as biomarkers.

Our hypothesis is that exo-miRNAs isolated from DIPG/DMG liquid biopsy could function as valid biomarkers, providing in real-time critical information on the therapeutic response.

The overall goal of the proposed project is to evaluate exo-miRNAs from DIPG/DMG liquid biopsy as surrogate biomarkers for disease diagnosis and therapeutic response.

Aim 1. To identify specific exosomal miRNA biomarkers of therapeutic response in the plasma of DIPG/DMG patients.

Aim 2. To evaluate the CSF derived exosomal miRNAs for pHGG molecular and/or locational subgrouping.

To achieve our aims, we will proceed in parallel on a prospective study and a retrospective study.

- The prospective study will take advantage of an ongoing clinical trial in our institutions (NCT03620032). The trial is a Phase 2, open label randomized study on the efficacy of two different radiotherapy schedules with the same concomitant and post-radiotherapy treatment based on nimotuzumab and vinorelbine, for newly diagnosed DIPG patients and an observational study for DMG with k27 mutation.

Blood samples will be collected from patients recruited on the trial between the Bambino Gesù Children’s Hospital (OPBG) and the Istituto Nazionale Tumori (INT). For standard and experimental arm of the trial, patients will be subjected to MRI evaluation every 12 weeks and for blood collection will be done concomitantly. Exo-miRNAs from patient plasma patients will be analysed to identify biomarkers of therapeutic response. Data will be integrated with MRI imaging analysis to determine the reliability of the identified biomarkers compared to standard neuro-radiological imaging.

- This part will be a retrospective study based on the analysis on CSF derived exo-miRNAs.

For this part we will count on a precious cohort of samples, including CSF and CSF-derived cell lines, from 15 patients. We will evaluate the CSF and CSF-derived cell lines to study the exo-miRNAs and their potential application for pHGG/DIPG subgrouping (as we have shown for plasma samples from 2018 funding). The CSF part of the study will represent the most challenging part due to the small volumes normally collected by lumbar puncture in children, but at the same time is highly innovative and worth it to be evaluated:

- CSF should represent a better reservoir of biomarkers (as closer to the tumour) when compared to the blood

- identified CSF exo-miRNAs will be compared with the plasma exo-miRNAs from same patients cohort.

We believe our proposed study may have a huge impact on the future clinical practice of DIPG by exploiting the exo-miRNAs from patient liquid biopsy as valuable non-invasive biomarkers.

The team of the PI and the collaborators have the required knowledge, expertise and access to institutional facilities to carry out the project.

Description of Research Proposal

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Budget

Lorem ipsum dolor sit amet, consectetur adipiscing elit. Integer gravida non felis non euismod. Fusce finibus aliquet consequat. Nam ac metus bibendum, iaculis purus sed, suscipit ligula. Proin et nisi libero. Mauris non urna urna. Nullam augue eros, fringilla sed mauris vitae, porta tincidunt risus. Aliquam sed tincidunt sem. Quisque lacinia quam tortor, imperdiet efficitur odio iaculis in. Sed ultricies condimentum volutpat. Vivamus dignissim faucibus porta.

Curabitur ut ipsum non odio malesuada vulputate. Morbi maximus, est eu lobortis molestie, tortor sapien hendrerit nisi, in cursus odio diam ut odio. Fusce pulvinar volutpat velit. Aliquam erat volutpat. Integer rhoncus mollis suscipit. Praesent non ipsum mollis, finibus nunc a, scelerisque nibh. In feugiat iaculis velit, eu semper lacus dignissim nec. Praesent vitae nisi leo. Cras venenatis dictum magna ut semper. Sed eget eros nibh. Sed vitae quam sed dolor faucibus elementum. Curabitur interdum porttitor finibus. Nullam tincidunt odio lectus, sit amet rhoncus libero dapibus sed. Sed mollis egestas enim, vel porta tortor volutpat eget.

Morbi orci urna, ornare non pretium eget, pulvinar eget magna. Ut consectetur efficitur varius. Fusce ac aliquet mauris, at mattis ligula. Quisque est libero, interdum id orci et, ornare luctus diam. Proin commodo lectus id accumsan blandit. Nulla eu turpis interdum, luctus ante ac, imperdiet tellus. In semper enim eu tristique aliquam.

Collaborations and Conflicts of Interest

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