Grants

DIPG remains a highly lethal disease with no effective treatment options. How DIPG cells reprogram nutrient consumption and usage via metabolic pathways to sustain their growth is not well understood. Using profiling of metabolites, we uncovered DIPG cells elevate the consumption and breakdown of branched chain amino acids (BCAAs); leucine, isoleucine and valine. Tumor cells also highly express the enzyme BCAT1, involved in breakdown of BCAAs. We believe that DIPG cells require BCAT1 to grow in cell culture. Our work aims to establish the reason why DIPG tumors depend on the metabolism of BCAAs, using methods to track the nutrients they are converted into, and by investigating why DIPG cells fail to grow when losing BCAT1 function. Second, we aim to test if there are therapeutic benefits for patients from reducing the levels of BCAAs in the diet through tailoring food composition. These diets have previously been established as safe and effective for treating specific metabolic disorders in children. Our proposed work will help understand how specific nutrients are utilized by DIPG cells and potentially present a novel way to eradicate DIPG growth and prolong survival of patients.

Aggressive brain cancers such as paediatric high-grade gliomas H3.3 and ATRX mutations are among the most challenging to treat, with limited options and poor outcomes. Recent discoveries, however, are opening promising new paths for targeted therapy.

Our research shows that mutations in key chromatin regulators—H3.3 and ATRX—not only drive tumour development but also create an unexpected vulnerability. These mutations disrupt regions of the genome called ribosomal DNA (rDNA), which produce RNA for building ribosomes—the cell’s protein factories. This disruption affects the nucleolus, a critical cell structure, and makes tumour cells unusually dependent on ribosome production.

This dependency offers a powerful opportunity for targeted treatment. We have shown that drugs blocking RNA Polymerase I (Pol I)—the enzyme responsible for rRNA synthesis—selectively impair tumour cells with these mutations while sparing healthy cells. Two such drugs, including a next-generation brain-penetrant compound, show strong promise in preclinical models.

Our project will uncover how H3.3 and ATRX mutations alter rDNA function and test Pol I inhibitors in advanced lab and animal models. By focusing on shared vulnerabilities, this work aims to deliver urgently needed, precision therapies for patients with currently untreatable brain cancers.

High-grade gliomas (HGG) are aggressive brain tumors that are a leading cause of cancer death in children and young adults. Some HGGs have specific changes in their DNA (like BRAF or NF1 mutations) that affect a pathway called RAS-ERK, which controls how cells grow and repair damage. While drugs that target this pathway (e.g., MEK inhibitors) can help, they don’t cure the cancer, and cancers often find ways to resist treatment.

Our research uses a different approach: combining MEK inhibitors with other treatments that damage the cancer cells' DNA. We discovered that when the RAS-ERK pathway is blocked, the tumor cells become worse at repairing DNA. This weakness makes them more vulnerable to treatments like chemotherapy or radiation. We are testing these combinations in lab-grown tumor cells and in mice with human brain tumors. Our goal is to find out which drug combinations work best and are safe enough to eventually try in children with HGG or similar tumors.

By learning how to use these weaknesses in cancer cells, we hope to create better, more effective treatments for young patients who currently have very few options.

Diffuse midline glioma (DMG) is a currently incurable childhood tumor that arises in the midline or brainstem of the brain. There are likely several reasons why treatments fail in these tumors; our research is focused on macrophages, which normally assist the body in fighting off disease and illness but are co-opted by the tumor to do the opposite. These tumor associated macrophages make up a large part of the tumor and can come from the blood or brain itself. In DMG, these macrophages appear to be switched “off” in a way that prevents them from attacking the tumor. Our research suggests that these cells play a key role in shutting down the body’s immune response and making the tumor resistant to treatment. We are focusing on genes and proteins expressed by these macrophages that are known to suppress the immune system. Using advanced lab models that mimic the human disease, we are testing whether blocking these proteins can help “wake up” the immune system to fight the tumor more effectively, especially when combined with radiation therapy. Ultimately, it is our mission to identify new therapeutic strategies targeting these macrophages to improve outcomes for children with this devastating disease.

Diffuse midline glioma (DMG) is a highly aggressive pediatric brain tumor with a dismal prognosis. A major treatment barrier is tumor hypoxia—low oxygen levels that reduce the effectiveness of radiotherapy and chemotherapy. Our recent research found that targeting mitochondrial metabolism, specifically by reducing oxygen consumption in tumor cells, can overcome hypoxia and improve treatment response. While the mitochondrial inhibitor phenformin showed promise, its clinical use is limited due to safety concerns, necessitating safer alternatives.

Through a high-throughput screen of FDA-approved drugs, we identified six candidates with mitochondrial inhibitory properties. Among them, berberine, sertindole, and penfluridol emerged as top candidates with strong anti-DMG effects. Berberine, used in traditional Chinese medicine, and the antipsychotic drugs sertindole and penfluridol all demonstrate excellent brain penetration and reduced tumor cell oxygen consumption in laboratory studies.

These drugs not only impair tumor metabolism but also slow tumor growth and enhance the effects of radiation. Our current work focuses on understanding their mechanisms of action, validating efficacy in preclinical models, and optimizing their use in combination with radiotherapy. This research aims to fast-track safer, more effective therapies for children with DMG and improve their currently grim outcomes.

Diffuse Intrinsic Pontine Glioma (DIPG) is an aggressive and fatal brain tumor that affects young children. Because it grows in the brainstem, it cannot be removed with surgery, and radiation offers only temporary relief. Chemotherapy and other standard treatments have not been effective. Most children with DIPG survive less than a year, highlighting the urgent need for new therapies.

One promising approach is CAR-T cell therapy, which uses engineered mmune cells, reprogrammed to recognize and kill cancer. However, DIPG has proven especially difficult to treat with CAR-T cells because the tumor creates a highly immunosuppressive environment. It actively shuts down immune cells and hides from them, preventing a sustained attack.

Our project uses advanced gene editing tools to protect CAR-T cells against these suppressive signals. By knocking out a specific combination of genes inside the T cells using a technique called base editing, we make them more resistant to suppression and cancer evasion. This allows the cells to survive longer, stay active, and more effectively eliminate tumor cells.

This research could lead to a new generation of immunotherapies that overcome the unique defenses of DIPG and offer real hope for children diagnosed with this devastating disease.

Diffuse Midline Gliomas or DMGs are a type of brain cancer that primarily arise in children and are incurable. This proposal will evaluate a novel therapy for DMG that targets an enzyme called LSD1.  We chose to focus on LSD1 as we have preliminary data that LSD1 turns off the immune system in DMG and so blocking LSD1 can activate the immune system against DMG.  We will evaluate a clinically relevant inhibitor to block LSD1, which is called bomedemstat, and is currently in late phase clinical trials in adults with cancer.  The investigative team is comprised of three laboratories with complementary expertise that has collaborated for years: the Becher lab has expertise in DMG biology and the development of mouse model systems to evaluate new therapies; the Chandra lab has a long track record of evaluating small molecules that block LSD1 in brain tumor models;  and the Bernstein lab has expertise in the basic biology of how proteins such as LSD1 work in cancer.  

Targeted radiopharmaceutical therapy (TRT) is an innovative form of radiation treatment. Unlike conventional external beam radiation therapy (EBRT), which irradiates both tumor and surrounding healthy tissue, TRT uses a radioactive drug that selectively binds tumor cells and delivers radiation directly at the site of disease. This principle has already shown remarkable success in adults with e.g., neuroendocrine tumors.

For children with diffuse intrinsic pontine glioma/diffuse midline glioma (DIPG/DMG) and medulloblastoma (MB), treatment options remain extremely limited, with radiotherapy only offering temporary relief and chemotherapy showing little benefit. There is an urgent need for novel, effective, and less toxic approaches.

With the generous support of The Cure Starts Now Foundation, our TARGET-FIRST project launched in 2022 to explore TRT as a potential therapy for pediatric brain tumors. In the first project phase, we identified and validated promising molecular targets, establishing a strong foundation for translational development (results presented at the 2025 Brain Cancer Symposium in Banff, Canada).

The next phase will test radiolabeled ligands for binding capacity and therapeutic efficacy in tumor cell lines and preclinical models. These studies will generate the first robust dataset for pediatric TRT, paving the way for precision-guided, less toxic treatments that could transform patient outcomes.

SUMOylation is a post-translational protein modification (PTM) involved in the control of gene regulation, DNA damage repair, immune responses, and tumor growth.  During the process, SUMO proteins are attached to target proteins by SUMOylation enzymes. Because of its importance in cancer, SUMOylation inhibitors are being developed as therapeutic agents. Subasumstat (TAK-981) is a first in-class SUMOylation inhibitor that disrupts SUMOylation. TAK-981 is now in multiple oncology clinical trials in patients with advanced cancers. Nothing is known about the importance of SUMOylation in DIPG. We discovered that SUMOylation genes are elevated in DIPG compared to normal brain and most other pediatric brain tumors. When tested in several DIPG cell lines, the SUMOylation inhibitor TAK-981 killed 70-90% DIPG cells in the Petri dish. In this grant we will test the efficacy of the drug in our mouse models of DIPG in combination with radiation. In addition to DIPG, SUMOylation genes are also elevated in two other pediatric brain tumors with poor outcomes - group 3 medulloblastoma and Atypical Teratoid Rhabdoid Tumor (ATRT). If the results of our studies in DIPG hold true in these two brain tumors, our research may have a broader impact in finding treatments for children with brain tumors.

In this project, we will bring a paradigm-shifting treatment approach, which has led to cures of other previously incurable childhood cancers, to diffuse midline glioma (DMG) for the first time: combination, multimodality therapy. Our clinical trial will incorporate radiation, locally delivered chemotherapy directly to the tumor, systemic chemotherapy to the whole brain and spinal cord, and medicine to harness the patient’s own immune system to fight their tumor. These treatments will be given in rational stages of therapy to maximize treatment effectiveness and patient safety. Our team brings extensive expertise and experience with DMG biology, clinical trial design, and each of the components of therapy. Our initial pilot trial is small, highly feasible, and achievable within a 12-18-month period. Most importantly, the trial will not be a static set of treatments. Instead, we will have input from many sources, including a committee of experts, a committee of family members, parallel work we will do in our labs, findings from other trials, and data from our own trial to help us continuously improve our trial treatment. Our overall goal is that this trial will improve over time until we achieve long-term survival for DMG patients with good quality of life.

We propose a new type of immunotherapy for diffuse midline glioma (DMG) and using gdT cells, a type of T cell that targets cancers differently than the T cells previously used in brain tumor immunotherapies, and using a novel immune-stimulating drug that we developed. Children with DMG need new therapies because radiation therapy and chemotherapy consistently fail to cure patients. Immunotherapy, which has cured previously incurable leukemias, is a promising new approach. Recently immunotherapies using T cells engineered to recognize DMG through Chimeric Antigen Receptors (CAR-T cells), successfully completed phase 1 trials. In these trials, CAR-T cells showed safety and striking effectiveness in individual patients. However, most DMG patients in these CAR-T trials suffered recurrence, suggesting that cure will likely require additional improvements to immunotherapy. To meet the need for improved immunotherapies, we propose to use gdT cells, which recognize and kill tumor cells even without CARs, to test new types of CAR-like innovations, and to add concurrent treatment with ResiPOx an immune stimulating agent that we formulated in nanoparticles to help it cross the blood brain barrier. We propose that these key innovations can bring about a step change and make immunotherapy for DMG newly effective.  

Pediatric high-grade gliomas are one of the most common causes of cancer-related death in children, and diffuse midline gliomas (DMGs) are the most rapidly fatal of these tumors. The infiltrative nature together with delicate tumor location in the brainstem precludes surgical resection, and the blood-brain barrier (BBB) prevents to deliver most drugs to the brain. To overcome this barrier, we will employ an innovative and effective delivery system, intranasal delivery (IND). IND is a practical, noninvasive method of bypassing the BBB to deliver drugs to the brain using unique anatomic (nerve) connection between nose to the brain, reducing unwanted systemic toxicity, and is amenable to convenient administration for patients. The novel altered cold-sore virus G207 has proven safe with evidence of responses and prolonged survival in children with high-grade gliomas. However, a key limitation of G207 is the use of intratumoral injection, which requires an invasive neurosurgical procedure and limits repeat injections, which would be beneficial for treating DMG and is made possible with IND. We will test the hypothesis that IND will facilitate giving repeated, maximum doses of G207 virus that can target DMG cells avoiding the need for invasive surgical procedures and without damaging healthy tissues.

High grade gliomas (HGG), including Diffuse Midline Gliomas (DMG), are highly aggressive brain cancers, with less than 5% survival at five years. DMGs are predominantly seen in children and young adults This proposal seeks to develop a novel immunotherapy to benefit these patients. Myeloid cells are white blood cells that are part of the immune system. We find that intravenous administration of immature myeloid cells lacking a protein called p50 (p50-IMC) slows the growth of HGG tumors in mouse models. p50-IMC develop into mature, activated tumor myeloid cells that direct T cells to attack the cancer. However, following p50-IMC therapy most tumors eventually progress. HGGs and DMGs express a sugar known as GD2 on their surface. To improve their efficacy, we propose to direct p50-IMC to HGG tumors by expressing a GD2-targeting chimeric antigen receptor (CAR) on the p50-IMC. We will determine the effectiveness of p50-IMC/GD2.CAR cells in a model of HGG. We will also evaluate the benefit of adding additional immunotherapy agents to GD2-targeted, activated myeloid cells, to assist them in stimulating T cells to attack the cancer cells. Upon completion of these studies, we anticipate evaluating our novel immunotherapy in clinical trials against HGGs and DMGs.

Aggressive brain tumors contain cancer stem cells that drive tumor aggressiveness and recurrence but remain elusive to detect. We have found that they acquire an abnormal sugar coating when they transition from a less aggressive to a more aggressive type. At Johns Hopkins, we have recently developed a specialized magnetic resonance imaging (MRI) technique that can detect these sugars which we hope to use as an imaging biomarker for the presence of glioma cancer stem cells in diffuse midline glioma (DMG).  Since this stem cell detection method does not require any injection of contrast agents it is easy and safe to implement with currently used MRI techniques. Building upon this innovation our ultimate goal is to develop and implement, from pre-clinical orthotopic mouse models to DMG patients, our new MRI technique to visualize high mannose levels in DMG stem cells. If succesful, this new imaging technique may then serve as a molecular imaging biomarker for predicting tumor aggressiveness and more importantly to diagnose tumor recurrence following surgery for further radiotherapy planning. Our first two scanned patients showed that mannose imaging is clinically feasible, with a never observed contrast that is distinct from that obtained with conventional MRI techniques currently.

Children diagnosed with diffuse midline glioma (DMG), a devastating brain cancer, face heartbreaking odds. Radiation, the only current treatment, provides limited benefit and causes serious, lifelong side effects. Our mission is to create safer, more effective therapies that give children a real chance to survive and thrive.

This project develops a new therapy using engineered immune cells, CAR T cells, designed to attack tumor cells while sparing normal cells. We are advancing a groundbreaking two-part strategy:

• Dual-target CAR therapy – We engineer a child’s immune cells to target two key molecules on tumor cells, reducing the chance of cancer escape and improving long-term effectiveness.
• Focused ultrasound – This non-invasive technology temporarily opens the brain’s protective barrier, enabling CAR T cells to reach tumors more efficiently and boosting their therapeutic activity.

Unlike current CAR T cells being tested in clinical trials, our preclinical studies use models that faithfully replicate both DMG tumor biology and the brain’s immune barriers. This enables rigorous evaluation of safety, delivery, and tumor-killing activity, generating critical data to accelerate the translation of a well-validated CAR therapy to first-in-child trials.