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123 abc lane, Townsville, ZZ 00000, USA
Funding Progress: $§ / $§§§§§
Epigenetic Regulation of Cell Fate Reprogramming in H3K27M-altered diffuse midline glioma
TranslationalDIPG, Childhood (Brain Cancer)
Lay Summary
Diffuse midline glioma, including those located at the pons in the brainstem (diffuse intrinsic pontine gliomas, DIPGs) are childhood brain tumors and patients with DIPG have a poor prognosis with a median survival time less than 1 year from diagnosis. Currently, apart from radiation therapy, which partially slows down tumor growth, there is no effective therapy for DIPG, and the patient outcome has not improved in the past decades. We have found that 5-azacytidine, a DNA demethylating agent, works effectively in animal models, inducing cell differentiation and improving survival. In this grant application, we request the funds to optimize the DNA demethylating agent, 5-azacytidine, in combination with histone deacetylase inhibitor, in animal models of DIPG. The drug combination reprograms the gene expression signature and is a clinical significance approach for DIPG patients. Furthermore, successful drug combinations will be tested with computed tomographic small animal ionized radiation in preclinical DIPG mouse models. If successful, we believe these findings would be the basis of actionable clinical hypothesis generation.
Executive Summary
Executive Summary
Pediatric diffuse midline gliomas (DMG), including those located at the pons in the brainstem (diffuse intrinsic pontine glioma, DIPG), are recognized as the most lethal childhood cancer. DIPG is a highly aggressive pediatric brain cancer that occurs in children of median age of 6-7 years. The median overall survival is around 12 months, and apart from radiation therapy that only transiently delays tumor growth, there is no effective therapy. Of note, patient survival has not changed in the past 50 years, despite intensive research. DMG is considered one of the most challenging problems in pediatric neuro-oncology, due to the lack of therapies and the tumor location that does not allow surgery resection. The most frequent genetic alteration in high-grade pediatric glioma is a somatic heterozygous mutation in histone 3 (H3) variants leading to lysine to methionine substitutions at position 27 (H3K27-altered). In particular, H3K27-altered accounts for about 80% of DIPG, while mutations in histone 3.3 G34R/V are more frequent in cerebral/cortical hemispheres gliomas. The putative molecular mechanism of H3K27M mutation is associated global loss of the repressive histone H3K27 trimethylated (H3K27me3) mark, inhibition of polycomb repressive complex 2 (PRC2), and significant changes in gene expression signature. It has been shown that epigenetic drugs can partially revert the global gene expression dysregulation and improve survival in preclinical models. In particular, the histone deacetylase inhibitor (HDACi), panobinostat, has been shown to cause growth reduction and improved survival in preclinical models. Clinical trials testing panobinostat in DIPG patients are ongoing to test drug safety. An additional epigenetic modulator that is often dysregulated in cancer is DNA methylation. DNA methylation is tightly controlled during the development to regulate the spatial-temporal gene induction and lineage commitment. In cancer, DNA demethylating agents have been used in cancer therapy and we have been shown that the DNA demethylating agent, 5-azacytidine (5-aza), induces cell differentiation and extends survival in glioma models. Since H3K27-altered DIPG has a subpopulation with stemness features and a blocked differentiation state, targeting the cell fate decision by epigenetic reprogramming the cancer cell might be an immediate therapy option. This rationale is based on the ability of panobinostat to reduce H3K27Ac density at super-enhancers, modulating enhancer and promoter regions of key genes, while 5-aza demethylates the DNA associated with developmental genes and tumor suppressor genes.
In this hypothesis-driven translational research project, our aim is to optimize a rationale epigenetic drug combination with targeted radiation therapy in the H3K27-altered DIPG mouse model and advance actionable clinical hypothesis generation. In aim 1 we will demonstrate the mechanism of 5-aza plus panobinostat in H3K27-altered DIPG cells. It has been shown that H3K27-altered DIPG has an aberrant gene expression with a stem cell gene expression signature and an undifferentiated state. In this aim, we will determine the effect of DNA demethylating agent (5-aza) in combination with HDACi (panobinostat) in the DNA methylation profile and gene expression signature in H3K27-altered DIPG cells. In addition, we will determine the biological effect of this epigenetic drug combination on cell growth (apoptosis, cell proliferation, and cell cycle), stem cell and glial differentiation markers, and human endogenous retrovirus expression (which is upregulated by 5-aza, Yamashita et al. unpublished data). In aim 2 we will determine the efficacy of 5-aza plus panobinostat in combination with targeted radiation therapy in intracranial DIPG models. Apart from radiation therapy, there are no valuable treatments for DIPG, and chemotherapy is ineffective for this cancer type. The compounds suggested to be evaluated in this research grant can target the infiltrative DMG reservoir, while radiation therapy can control the tumor core. Our short-term goal is better understand the biology of H3K27-altered DIPG and how to implement the best therapy combination in preclinical models. Our long-term goal is to improve the quality of life and extend survival in DIPG patients.
Our research plan is designed to determine if and how to best use these epigenetic drugs in intracranial DIPG mouse models. Drug administration and dose, intracranial tumor implantation, and molecular biology experiments are established in our laboratory, providing compelling evidence of study feasibility. We hope to demonstrate that epigenetic drugs substantially enhance the survival in animal models with the standard of care radiation. The results from this work will allow us to design better and possibly support a new trial for patients with H3K27-altered glioma, which account for a substantial fraction of brain cancer mortality. Finally, our lab serves as a training center for students and post-doctoral fellows with an established pipeline for trainees with no commercial interest. We are supported for the near future almost entirely by peer-reviewed grants and rely on competitive external funding for our dual mission of education and clinical advances.
Description of Research Proposal
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Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Donec faucibus, turpis sit amet maximus dapibus, sapien nisl bibendum turpis, pharetra commodo tellus libero vel nulla. Sed nec velit viverra, congue sapien et, gravida libero. Proin eget ante eget turpis egestas accumsan. Aliquam arcu nibh, aliquam rhoncus vulputate in, pellentesque at sem. Maecenas cursus tempus nibh id tempus. Mauris dolor sapien, lacinia sit amet condimentum at, dapibus eget lectus. Phasellus vel pellentesque ex. Nunc aliquam in ligula at tristique. In mollis suscipit felis eu finibus. Nullam non dignissim nibh, nec suscipit ex. Suspendisse tincidunt et mauris id finibus. Aliquam vehicula a sem quis venenatis.
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Pellentesque nibh erat, egestas sit amet sagittis malesuada, rhoncus at neque. Mauris maximus commodo tortor, non egestas magna finibus vitae. Sed hendrerit nulla vel venenatis tempor. Suspendisse tristique tincidunt libero et placerat. Nam tincidunt condimentum lorem, vel pharetra est iaculis sed. Nullam ac tincidunt orci. Quisque pharetra ut sem sit amet aliquam. Phasellus risus libero, varius in condimentum vel, commodo id ipsum. Aliquam in metus cursus, mattis diam ut, aliquam magna. Suspendisse facilisis dui et orci varius, suscipit facilisis augue dapibus. In eget nibh ipsum. Suspendisse eget pharetra est, quis condimentum felis. Fusce scelerisque congue libero, sed aliquam mi elementum a. Etiam scelerisque ante non auctor porta. Nam eu nunc id ex finibus dictum. Praesent dui ex, dictum ac massa eget, rutrum gravida nisi.
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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
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.