Primary Brain Research Articles

CBTRUS Statistical Report: Primary Brain and Other Central Nervous System Tumors Diagnosed in the United States in 2017-2021.

The Central Brain Tumor Registry of the United States (CBTRUS), in collaboration with the Centers for Disease Control and Prevention and the National Cancer Institute, is the largest population-based registry focused exclusively on primary brain and other central nervous system (CNS) tumors in the United States (US) and represents the entire US population. This report contains the most up-to-date population-based data on primary brain tumors available and supersedes all previous reports in terms of completeness and accuracy. All rates are age-adjusted using the 2000 US standard population and presented per 100,000 population. Between 2017 and 2021, the average annual age-adjusted incidence rate (AAAIR) of all primary malignant and non-malignant brain and other CNS tumors was 25.34 per 100,000 population (malignant AAAIR=6.89 and non-malignant AAAIR=18.46). This overall rate was higher in females compared to males (28.77 versus 21.78 per 100,000) and non-Hispanic Black persons compared to persons who were non-Hispanic White (26.60 versus 25.72 per 100,000), non-Hispanic American Indian/Alaska Native (23.48 per 100,000), non-Hispanic Asian or Pacific Islander (19.86 per 100,000), and Hispanic persons of all races (22.37 per 100,000). Gliomas accounted for 22.9% of all tumors. The most commonly occurring malignant brain and other CNS histopathology was glioblastoma (13.9% of all tumors and 51.5% of all malignant tumors), and the most common predominantly non-malignant histopathology was meningioma (41.7% of all tumors and 56.8% of all non-malignant tumors). Glioblastomas were more common in males, and meningiomas were more common in females. In children and adolescents (ages 0-19 years), the incidence rate of all primary brain and other CNS tumors was 6.02 per 100,000 population. There were 87,053 deaths attributed to malignant brain and other CNS tumors between 2017 and 2021. This represents an average annual mortality rate of 4.41 per 100,000 population and an average of 17,411 deaths per year. The five-year relative survival rate following diagnosis of a malignant brain or other CNS tumor was 35.7%. For a non-malignant brain or other CNS tumor the five-year relative survival rate was 92.0%.

Molecular Determinants for Photodynamic Therapy Resistance and Improved Photosensitizer Delivery in Glioma.

Gliomas account for 24% of all the primary brain and Central Nervous System (CNS) tumors. These tumors are diverse in cellular origin, genetic profile, and morphology but collectively have one of the most dismal prognoses of all cancers. Work is constantly underway to discover a new effective form of glioma therapy. Photodynamic therapy (PDT) may be one of them. It involves the local or systemic application of a photosensitive compound-a photosensitizer (PS)-which accumulates in the affected tissues. Photosensitizer molecules absorb light of the appropriate wavelength, initiating the activation processes leading to the formation of reactive oxygen species and the selective destruction of inappropriate cells. Research focusing on the effective use of PDT in glioma therapy is already underway with promising results. In our work, we provide detailed insights into the molecular changes in glioma after photodynamic therapy. We describe a number of molecules that may contribute to the resistance of glioma cells to PDT, such as the adenosine triphosphate (ATP)-binding cassette efflux transporter G2, glutathione, ferrochelatase, heme oxygenase, and hypoxia-inducible factor 1. We identify molecular targets that can be used to improve the photosensitizer delivery to glioma cells, such as the epithelial growth factor receptor, neuropilin-1, low-density lipoprotein receptor, and neuropeptide Y receptors. We note that PDT can increase the expression of some molecules that reduce the effectiveness of therapy, such as Vascular endothelial growth factor (VEGF), glutamate, and nitric oxide. However, the scientific literature lacks clear data on the effects of PDT on many of the molecules described, and the available reports are often contradictory. In our work, we highlight the gaps in this knowledge and point to directions for further research that may enhance the efficacy of PDT in the treatment of glioma.

Inter-tissue differences in oxidative stress susceptibility reveal a less stable endothelial barrier in the brain than in the retina

Oxidative stress can impair the endothelial barrier and thereby enable autoantibody migration in Neuromyelitis optica spectrum disorder (NMOSD). Tissue-specific vulnerability to autoantibody-mediated damage could be explained by a differential, tissue-dependent endothelial susceptibility to oxidative stress. In this study, we aim to investigate the barrier integrity and complement profiles of brain and retinal endothelial cells under oxygen-induced oxidative stress to address the question of whether the pathomechanism of NMOSD preferentially affects the brain or the retina.Primary human brain microvascular endothelial cells (HBMEC) and primary human retinal endothelial cells (HREC) were cultivated at different cell densities (2.5*104 to 2*105 cells/cm2) for real-time cell analysis. Both cell types were exposed to 100, 500 and 2500 μM H2O2. Immunostaining (CD31, VE-cadherin, ZO-1) and Western blot, as well as complement protein secretion using multiplex ELISA were performed.HBMEC and HREC cell growth phases were cell type-specific. While HBMEC cell growth could be categorized into an initial peak, proliferation phase, plateau phase, and barrier breakdown phase, HREC showed no proliferation phase, but entered the plateau phase immediately after an initial peak. The plateau phase was 7 h shorter in HREC. Both cell types displayed a short-term, dose-dependent adaptive response to H2O2. Remarkably, at 100 μM H2O2, the transcellular resistance of HBMEC exceeded that of untreated cells. 500 μM H2O2 exerted a more disruptive effect on the HBMEC transcellular resistance than on HREC. Both cell types secreted complement factors H (FH) and I (FI), with FH secretion remaining stable after 2 h, but FI secretion decreasing at higher H2O2 concentrations.The observed differences in resistance to oxidative stress between primary brain and retinal endothelial cells may have implications for further studies of NMOSD and other autoimmune diseases affecting the eye and brain. These findings may open novel perspectives for the understanding and treatment of such diseases.

Ultra-low-field magnetic resonance angiography at 0.05 T: A preliminary study.

We aim to explore the feasibility of head and neck time-of-flight (TOF) magnetic resonance angiography (MRA) at ultra-low-field (ULF). TOF MRA was conducted on a highly simplified 0.05 T MRI scanner with no radiofrequency (RF) and magnetic shielding. A flow-compensated three-dimensional (3D)gradient echo (GRE) sequence with a tilt-optimized nonsaturated excitation RF pulse, and a flow-compensated multislice two-dimensional (2D) GRE sequence, were implemented for cerebral artery and vein imaging, respectively. For carotid artery and jugular vein imaging, flow-compensated 2D GRE sequences were utilized with venous and arterial blood presaturation, respectively. MRA was performed on young healthy subjects. Vessel-to-background contrast was experimentally observed with strong blood inflow effect and background tissue suppression. The large primary cerebral arteries and veins, carotid arteries, jugular veins, and artery bifurcations could be identified in both raw GRE images and maximum intensity projections. The primary brain and neck arteries were found to be reproducible among multiple examination sessions. These preliminary experimental results demonstrated the possibility of artery TOF MRA on low-cost 0.05 T scanners for the first time, despite the extremely low MR signal. We expect to improve the quality of ULF TOF MRA in the near future through sequence development and optimization, ongoing advances in ULF hardware and image formation, and the use of vascular T1 contrast agents.

Abstract CT178: Phase 1 trial of PF-07799933 (ARRY-440), a next-generation BRAF inhibitor, for BRAF-mutant cancers

Abstract BACKGROUND Approved BRAF inhibitors (BRAFi) have transformed the treatment of BRAF V600-mutant cancers. However, clinical benefit is limited by BRAF dimer-promoting resistance mutations, toxicity from paradoxical activation of signaling in BRAF wild-type cells, and poor brain penetration. Further, current RAF inhibitors are ineffective against non-V600 BRAF mutants. Next-generation agents that cause pan-RAF inhibition may more broadly target BRAF mutants but are limited by a narrow therapeutic index. PF-07799933 (ARRY-440) is a brain-penetrant BRAF-selective monomer/dimer inhibitor that spares ARAF and CRAF. METHODS We characterized PF-07799933 in patient (pt)-derived BRAF-mutant cancer cells in vitro and in vivo. We investigated PF-07799933 in pts with refractory BRAF-mutant solid tumors using a novel first-in-patient phase 1 design that enabled flexible, rapid dose escalation based on safety and pharmacokinetics (PK) assessments. RESULTS PF-07799933 inhibited signaling in vitro, disrupted BRAF-containing homo- and heterodimers, and caused less paradoxical signaling than approved agents. PF-07799933 alone and in combination with binimetinib inhibited tumor growth systemically and in the brain in mouse xenografts harboring de novo and acquired BRAF dimer-forming mutations. As of August 24, 2023 data cutoff, 30 pts with BRAF-mutant cancers started PF-07799933 treatment at 50 mg QD-450 mg BID as monotherapy (60%) or combination with binimetinib (27%) or cetuximab (all colorectal cancer [CRC]) (13%). Tumor types were melanoma (mel) (43%), CRC (17%), primary brain (PBT) (13%), thyroid (10%), and other (n=1/3% each). BRAF mutations were class I (V600E) (73%), Class II (13%), and class III (13%). All BRAF V600E+ cancer pts were previously treated with ≥1 approved BRAFi and all BRAF V600E+ mel pts also were previously treated with ≥1 immune checkpoint inhibitor. PF-07799933 was well-tolerated as monotherapy or combination. There were no DLTs and the MTD was not reached. Treatment-emergent adverse events (TEAEs) in ≥3 pts for monotherapy were fatigue (any grade 44%/grade ≥3 0%), headache (28%/0%), vision blurred (22%/6%), and lipase increased (16%/0%). The most common TEAEs for combination were peripheral edema (33%/0%), acneiform rash, diarrhea, and fatigue (each 28%/0%). The AE profile was consistent with less paradoxical signaling activation. PK-guided dose escalation enabled PF-07799933 efficacious concentrations to be reached by dose level 3 (225 mg BID), together with multiple confirmed responses in BRAF V600E+ cancer pts (n=4/7 at 225 mg BID ± binimetinib, includes 1 complete response [CR] in a BRAF V600E+ PBT pt) systemically, in the brain, and all after failure of approved BRAF inhibitors. A 5th confirmed response (2nd BRAF V600E+ PBT pt) occurred after data cutoff. This includes, to our knowledge, the first evidence of efficacy in a BRAF V600E+ pt with a dimerizing BRAF splice variant. A BRAF G466E+ (class III) adenoid cystic carcinoma pt had a molecular CR in ctDNA. CONCLUSION PF-07799933 treatment was associated with anti-tumor activity against BRAF V600- and non-V600-mutant cancers preclinically and in treatment-refractory pts. The novel, rapid PK-informed dose escalation design provides a new paradigm for accelerating testing of next-generation targeted therapies early in clinical development. TRIAL REGISTRATION NUMBER NCT05355701 Citation Format: Rona Yaeger, Meredith McKean, Rizwan Haq, Joseph T. Beck, Matthew H. Taylor, Jonathan Cohen, Daniel Bowles, Shirish M. Gadgeel, Catalin Mihalcioiu, Kyriakos P. Papadopoulos, Eli L. Diamond, Keren B. Sturtz, Gang Feng, Stefanie K. Drescher, Micaela B. Reddy, Bhaswati Sengupta, Arnab K. Maity, Suzy A. Brown, Anurag Singh, Eric N. Brown, Brian R. Baer, Jim Wong, Tung-Chung Mou, Wen I. Wu, Dean Kahn, Sunyana Gadal, Neal Rosen, John J. Gaudino, Patrice A. Lee, Dylan P. Hartley, Stephen Michael Rothenberg. Phase 1 trial of PF-07799933 (ARRY-440), a next-generation BRAF inhibitor, for BRAF-mutant cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr CT178.

CBTRUS Statistical Report: Primary Brain and Other Central Nervous System Tumors Diagnosed in the United States in 2016-2020.

The Central Brain Tumor Registry of the United States (CBTRUS), in collaboration with the Centers for Disease Control and Prevention and the National Cancer Institute, is the largest population-based registry focused exclusively on primary brain and other central nervous system (CNS) tumors in the United States (US) and represents the entire US population. This report contains the most up-to-date population-based data on primary brain tumors available and supersedes all previous CBTRUS reports in terms of completeness and accuracy. All rates are age-adjusted using the 2000 US standard population and presented per 100,000 population. The average annual age-adjusted incidence rate (AAAIR) of all malignant and non-malignant brain and other CNS tumors was 24.83 per 100,000 population (malignant AAAIR=6.94 and non-malignant AAAIR=17.88). This overall rate was higher in females compared to males (27.85 versus 21.62 per 100,000) and non-Hispanic persons compared to Hispanic persons (25.24 versus 22.61 per 100,000). Gliomas accounted for 26.3% of all tumors. The most commonly occurring malignant brain and other CNS histopathology was glioblastoma (14.2% of all tumors and 50.9% of all malignant tumors), and the most common predominantly non-malignant histopathology was meningioma (40.8% of all tumors and 56.2% of all non-malignant tumors). Glioblastomas were more common in males, and meningiomas were more common in females. In children and adolescents (ages 0-19 years), the incidence rate of all primary brain and other CNS tumors was 6.13 per 100,000 population. There were 86,030 deaths attributed to malignant brain and other CNS tumors between 2016 and 2020. This represents an average annual mortality rate of 4.42 per 100,000 population and an average of 17,206 deaths per year. The five-year relative survival rate following diagnosis of a malignant brain and other CNS tumor was 35.7%, for a non-malignant brain and other CNS tumor the five-year relative survival rate was 91.8%.

Delay in Time to Oncologic Therapies for Patients with Positive COVID-19 Test

For several cancers, delays between diagnosis and initiation of treatment has important clinical implications, often affecting trial eligibility, treatment intention, and oncologic outcomes. The coronavirus disease-19 (COVID-19) pandemic placed an extraordinary strain on the United States healthcare system, and its effect on oncologic patterns of care has yet to be established. We hypothesize that patients who received a new cancer diagnosis and subsequently tested positive for COVID-19 had delayed oncologic treatment compared to those who did not test positive for COVID-19. The National Cancer Database (NCDB) was queried to identify patients who were diagnosed and treated for any of 10 common malignancies from 2019-2020. Included disease sites were head and neck, esophagus, rectum, anus, lung, breast, cervix, uterus, prostate, and primary brain. Those who tested positive for COVID-19 between time of diagnosis and first oncologic treatment (including surgery, radiation, or systemic therapy) were compared to those who did not test positive for COVID-19. COVID-19 positivity was assessed using a new variable in the NCDB, "SARSCOV2_POS," which captures whether patients received a positive COVID-19 test via reverse transcriptase-polymerase chain reaction testing in inpatient, outpatient, or emergency room settings in 2020. Duration in days from cancer diagnosis to time to first treatment (TTFT) was analyzed using two-sample t-tests, with significance level of p<0.05. A total of 1,503,127 patients were identified for analysis. Of these, 7,340 (0.5%) tested positive for COVID-19 between diagnosis and start of treatment. Initial treatment was most commonly surgery (55.3%), followed by systemic therapy (17.4%) and radiation (12.7%). Overall, median TTFT was 55 days [interquartile range (IQR) 31-91] for the COVID-19 group versus 34 days (IQR 15-56) for the non-COVID-19 group (p <0.01). Subgroup analysis of the 10 individual sites of disease revealed statistically significant delays in each, with greatest absolute difference in median TTFT in prostate (31.5 days; 95.5 versus 64.0) and greatest relative difference in brain (>700%, 28.5 versus 4.0). In the first year of the pandemic, patients who tested positive for COVID-19 between oncologic diagnosis and initial management experienced significant delays in initiation of cancer-directed therapy compared to those who did not test positive for COVID-19, with median increase in TTFT of 21 days. Additional follow-up is needed to evaluate the clinical impact of these delays, as well as change in patterns of care in later years of the pandemic.

Nanoparticle Strategies to Improve the Delivery of Anticancer Drugs across the Blood-Brain Barrier to Treat Brain Tumors.

Primary brain and central nervous system (CNS) tumors are a diverse group of neoplasms that occur within the brain and spinal cord. Although significant advances in our understanding of the intricate biological underpinnings of CNS neoplasm tumorigenesis and progression have been made, the translation of these discoveries into effective therapies has been stymied by the unique challenges presented by these tumors' exquisitely sensitive location and the body's own defense mechanisms (e.g., the brain-CSF barrier and blood-brain barrier), which normally protect the CNS from toxic insult. These barriers effectively prevent the delivery of therapeutics to the site of disease. To overcome these obstacles, new methods for therapeutic delivery are being developed, with one such approach being the utilization of nanoparticles. Here, we will cover the current state of the field with a particular focus on the challenges posed by the BBB, the different nanoparticle classes which are under development for targeted CNS tumor therapeutics delivery, and strategies which have been developed to bypass the BBB and enable effective therapeutics delivery to the site of disease.

Mivavotinib, a Syk Inhibitor, in Relapsed/Refractory (R/R) Non-GCB Diffuse Large B-Cell Lymphoma (DLBCL) with or without MYD88 and/or CD79 Mutations: A Phase 2 Study

Background: Spleen tyrosine kinase (Syk) is a key signaling protein that activates survival pathways in B-cell lymphomas. The non-germinal center B-cell like (non-GCB) or activated B-cell like (ABC) cell-of-origin subtype of DLBCL has greater reliance on the B-cell receptor pathway and Syk signaling for survival, compared with the GCB subtype. Multiple signaling pathways are activated by Syk in ABC DLBCL (NF-κB and phosphoinositide 3-kinase [PI3K]), whereas only the PI3K pathway is activated by Syk in GCB DLBCL. MYD88 and/or CD79b mutations are strongly enriched within ABC DLBCL and are associated with poor prognosis. Syk hyperactivation has been observed in ABC DLBCL cell lines harboring MYD88/CD79 mutations, with activation of NF-κB via Syk-mediated signaling through both the BCR and the toll-like-receptor (TLR)-MYD88 pathways. Mivavotinib is an orally dosed, potent, and selective Syk inhibitor, which showed potent single agent activity in preclinical studies in ABC and GCB DLBCL cell lines, with the strongest activity against those harboring MYD88 and/or CD79b mutations. In the clinical setting, heavily pre-treated all-comer R/R DLBCL patients receiving single agent mivavotinib at the recommended phase 2 dose (100 mg once daily [QD]) showed an overall response rate (ORR) of 33% (16% complete responses [CR]) with durable responses (median duration of response [DoR] not reached [95% CI: 7 months - NE]) and a favorable safety profile. Subgroups by cell-of-origin showed a substantially higher ORR in patients with the non-GCB subtype (53%; 95% CI: 27, 79) than GCB (22%; 95% CI: 0.11, 0.37), and a median DoR of 15.7 months (95% CI: 2.2, NE) in non-GCB responders. CX-659-401 (NCT05319028) is an ongoing study evaluating two dosing regimens of mivavotinib monotherapy in patients with R/R non-GCB DLBCL, in whom circulating tumor DNA-based next generation sequencing (NGS) will be used to determine MYD88/CD79b mutational status. The goals of this strategy are to confirm the activity of mivavotinib previously observed in non-GCB DLBCL, evaluate its activity in DLBCL according to mutational status of MyD88/CD79b, and optimize the dose and schedule in this biomarker-defined patient population. Methods: This multicenter, open-label, phase 2 study will enroll patients with histologically confirmed de novo or transformed DLBCL that is non-GCB by immunohistochemistry (IHC) per Hans algorithm or ABC by gene expression profiling, including primary mediastinal and primary extranodal DLBCL. Patients must have had at least 2 but no more than 5 prior lines of systemic anticancer therapy. Following standard first-line R-CHOP (or equivalent), patients must have received 2nd-line salvage therapy with or without autologous stem cell transplant (ASCT), and/or CAR-T therapy, unless ineligible for 2nd-line salvage, ASCT, or CAR-T. Additional key eligibility criteria include age >18 years, ECOG performance status 0-2, and FDG-PET-avid measurable disease per 2014 Lugano criteria. DLBCL patients with primary brain or secondary central nervous system involvement with active disease will be excluded. Patients will be randomized to 1 of 2 arms (n=25 planned per arm): (1) 100 mg QD; or (2) 120 mg QD for 2 weeks followed by 80 mg QD thereafter; in 28-day cycles until disease progression or unacceptable toxicity. The primary endpoints are ORR (by independent radiology review according to 2014 Lugano Criteria) and safety/tolerability. Secondary endpoints include DoR, progression-free survival (PFS), overall survival (OS), and CR rate. Exploratory endpoints include pharmacokinetic, biomarker assessments, and investigator-assessed ORR, DoR, PFS, and CR rate. Results of this study will help to define the optimal dose and provide confirmatory efficacy and safety data of mivavotinib in patients with non-GCB DLBCL, including MYD88/CD79b-mutated DLBCL to guide further expansion in these biomarker-defined cohorts.

NCOG-40. UTILITY OF THE SEIZURE CONTROL COMPOSITE INDEX (RANO-SCCI) IN EVALUATING SEIZURES IN CNS TUMOR PATIENTS

Abstract BACKGROUND Seizures are a common symptom in patients with CNS tumors. The RANO-Seizure Control Composite Index (RANO-SCCI) was developed to monitor seizures over time by patients and clinicians. We describe the use of the RANO-SCCI as part of the NCI NOB-Natural History Study. METHODS Patients completed a RANO-SCCI prior to their clinic appointment with results shared with clinicians who then completed a RANO-SCCI as part of the same appointment. Seizure reports were tallied by respondent type and further by tumor type and grade. Concordance between patient and clinician report was also investigated. RESULTS One hundred fifty-seven patients underwent 250 clinical evaluations. Participants were primarily white (82%) males (57%) with median age 47 years (range: 19 – 78). Most had high grade (60%) primary brain (86%) tumors, were in surveillance (76%), with no prior recurrence (57%) and KPS ≥ 90 (76%). Half of patients and 62% of clinicians reported the patient ever having a seizure. The 12% discordance were all where the patient did not report a seizure, but clinicians did. There was good agreement on seizure reporting since last visit (74% patients, 78% clinicians), and anti-seizure medication use (51% patients, 57% clinicians). Seizures were reported more frequently in patients with high-grade tumors (WHO Grade 3 or 4) (60% patients, 76% clinicians) versus low-grade (WHO Grade 1 or 2) (51% patients, 59% clinicians), and among patients with oligodendrogliomas (71% patients, 85% clinicians) versus non-oligodendroglioma tumors (54% patients, 68% clinicians). CONCLUSIONS The RANO-SCCI was used for self-report and clinician report of seizure activity. A higher frequency was reported among oligodendroglioma and high-grade tumors. Disparities between patient and clinician responses may reveal a limitation in the patient’s ability to recognize and assess their seizure symptoms. Further work will investigate reporting congruence and associations with clinical and demographic characteristics.

CBTRUS Statistical Report: Primary Brain and Other Central Nervous System Tumors Diagnosed in the United States in 2015-2019.

The Central Brain Tumor Registry of the United States (CBTRUS), in collaboration with the Centers for Disease Control and Prevention and the National Cancer Institute, is the largest population-based registry focused exclusively on primary brain and other central nervous system (CNS) tumors in the United States (US) and represents the entire US population. This report contains the most up-to-date population-based data on primary brain tumors available and supersedes all previous reports in terms of completeness and accuracy. All rates are age-adjusted using the 2000 US standard population and presented per 100,000 population. The average annual age-adjusted incidence rate (AAAIR) of all malignant and non-malignant brain and other CNS tumors was 24.71 per 100,000 population (malignant AAAIR=7.02 and non-malignant AAAIR=17.69). This overall rate was higher in females compared to males (27.62 versus 21.60 per 100,000) and non-Hispanic persons compared to Hispanic persons (25.09 versus 22.95 per 100,000). The most commonly occurring malignant brain and other CNS histopathology was glioblastoma (14.2% of all tumors and 50.1% of all malignant tumors), and the most common non-malignant histopathology was meningioma (39.7% of all tumors and 55.4% of all non-malignant tumors). Glioblastoma was more common in males, and meningiomas were more common in females. In children and adolescents (ages 0-19years), the incidence rate of all primary brain and other CNS tumors was 6.20 per 100,000 population. An estimated 93,470 new cases of malignant and non-malignant brain and other CNS tumors are expected to be diagnosed in the US population in 2022 (26,670 malignant and 66,806 non-malignant). There were 84,264 deaths attributed to malignant brain and other CNS tumors between 2015 and 2019. This represents an average annual mortality rate of 4.41 per 100,000 population and an average of 16,853 deaths per year. The five-year relative survival rate following diagnosis of a malignant brain and other CNS tumor was 35.7%, while for non-malignant brain and other CNS tumors the five-year relative survival rate was 91.8%.

Combined MEK and JAK/STAT3 pathway inhibition effectively decreases SHH medulloblastoma tumor progression

Medulloblastoma (MB) is the most common primary malignant pediatric brain cancer. We recently identified novel roles for the MEK/MAPK pathway in regulating human Sonic Hedgehog (SHH) MB tumorigenesis. The MEK inhibitor, selumetinib, decreased SHH MB growth while extending survival in mouse models. However, the treated mice ultimately succumbed to disease progression. Here, we perform RNA sequencing on selumetinib-treated orthotopic xenografts to identify molecular pathways that compensate for MEK inhibition specifically in vivo. Notably, the JAK/STAT3 pathway exhibits increased activation in selumetinib-treated tumors. The combination of selumetinib and the JAK/STAT3 pathway inhibitor, pacritinib, further reduces growth in two xenograft models and also enhances survival. Multiplex spatial profiling of proteins in drug-treated xenografts reveals shifted molecular dependencies and compensatory changes following combination drug treatment. Our study warrants further investigation into MEK and JAK/STAT3 inhibition as a novel combinatory therapeutic strategy for SHH MB.

Maslinic Acid Inhibits the Growth of Malignant Gliomas by Inducing Apoptosis via MAPK Signaling.

Background Gliomas are primary malignant brain tumors. Despite recent advances in surgery and clinical neuro-oncology, the prognosis of patients with glioma is still poor. Therefore, there is an urgent need to find new therapeutic drugs. Methods Here, we have studied the anticancer effect of maslinic acid in glioma and explored its potential molecular mechanism. CCK-8, Ki67 immunofluorescence, and colony formation tests are used to detect the proliferation of glioma cells. Transwell and migration experiments are used to detect the function of cell invasion and migration, and RNA-seq was performed to identify differentially expressed genes. Western blot analysis helps us identify important signaling pathways. Finally, the anticancer effect of maslinic acid was confirmed in vivo through tumor xenografting experiments. Results Our experiments obtained high-throughput data on the treatment of maslinic acid in glioma. We found that maslinic acid significantly inhibits the proliferation, invasion, and migration of glioma cells and promotes the apoptosis of glioma cells via suppressing MAPK signaling. Conclusions This is the first time to analyze the mechanism of maslinic acid against glioma based on transcription. Our experiments show that maslinic acid may be a useful natural product for the treatment of glioma.

Abstract 6296: Preclinical evaluation of a panel of FGFR inhibitors for their normal brain and brain tumor distribution

Abstract Glioblastoma (GBM) is the most prevalent and lethal primary intrinsic brain tumor in adults with approximately 5-8% of GBM patients displaying fibroblast growth factor receptor (FGFR) gene mutations. This subset of GBM patients could potentially benefit from targeted FGFR inhibitor treatment, thus making it imperative to explore FGFR inhibitors as a therapeutic option. In this study, seven pan-FGFR inhibitors (infigratinib, AZD4547, debio1347, erdafitinib, LY2874455, pemigatinib, and TAS-120) were evaluated for their brain penetration ability employing cassette dosing strategy with an aim to identify the optimal candidates for brain tumor targeting. Cassette-dosing strategy was confirmed to be an acceptable approach for screening the panel of FGFR inhibitors by comparing pharmacokinetic parameters obtained from discrete- vs cassette-dosing studies. Subsequently, cassette-dosing strategy was utilized to investigate distributional kinetics of FGFR inhibitors in normal brain and brain tumor in wild-type and triple-knockout (Mdr1a/b-/-Bcrp1-/-) mice. The comparison of brain-to-plasma partition coefficients for total (Kp,brain) and unbound (Kp,brain) drug levels in wild-type and triple-knockout mice revealed the importance of major blood-brain barrier (BBB) efflux transporters in four out of seven FGFR inhibitors. The impact of BBB and efflux transporters were further evaluated in flank and orthotopic patient-derived xenograft mouse models of brain tumors. All FGFR inhibitors were shown to accumulate significantly in both flank and orthotopic brain tumors relative to normal brain. Furthermore, the levels in flank brain tumors were at least 10-fold higher as compared to the levels in orthotopic brain tumors with five FGFR inhibitors, indicating the critical role of efflux transporters and the impact of BBB disruption in the brain and brain tumor distribution. The predictive parameters of brain penetration calculated based on physico-chemical properties have been estimated and shown not to correlate with the experimentally obtained neuro-pharmacokinetic values. In summary, our study provides comprehensive evaluation of neuro-pharmacokinetic behavior of seven FGFR inhibitors and delivers rationale for selection of most optimal candidates for future investigation in brain tumor clinical trials. Citation Format: William Knight, Kamal Shaik, Tigran Margaryan, Shwetal Mehta, Nader Sanai, Artak Tovmasyan. Preclinical evaluation of a panel of FGFR inhibitors for their normal brain and brain tumor distribution [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6296.

Abstract 5983: Gene profile analysis of breast cancer lung metastasis, liver metastasis, brain metastasis and primary tumor

Abstract Background: The incidence of male breast cancer (MBC) is lower than female breast cancer (FBC), representing 1% of all breast cancer worldwide. The difference with FBC may affect the treatment strategy of MBC, but the necessary precondition is that we should know more about the difference between FBC and MBC. However, Because of its low incidence that the relevant data about MBC is relatively scarce. Here we report some data about gene mutation of MBC, and compared it with FBC. Methods: Blood or tumor Tissue samples of breast cancer were collected and subjected to NGS in a College of American Pathologists (CAP)-certified and Clinical Laboratory Improvement Amendments (CLIA)-accredited lab for gene mutation analysis (3D Medicines Inc.,Shanghai, China). Statistical analysis was performed using GraphPad Prism (version 7.01) and SPSS version 21.0 (SPSS,Inc.). Results: Approximately 182 samples were analyzed, among which, exist 82 samples were from primary tissue, 66 samples were breast cancer liver metastases, 20 were lung metastases and 14 were brain metastases. High genomic heterogeneity was found among primary breast cancer tissue, breast cancer liver metastases, lung metastases, brain metastases, that is at least 20 gene mutations were found in these BC samples, while different gene mutations were showed among primary and different metastases tissues. The top five mutated gene in primary tumor tissue were TP53(80.49%), PIK3CA(53.66%), FGF19(30.49%), ERBB2(26.83%), MYC(23.17%); in liver metastases tissues were TP53(54.55%), FGF19(39.39%), PIK3CA(37.88%), ERBB2(21.21%), CCND1(19.70%); in lung metastases tissues were TP53(70.00%), PIK3CA(55.00%), ERBB2,MCL1(25.00%), CDKN2A/2B(20.00%), CCND1, NF1, CDK12(15.00%), CCND2, MYC, PTEN, GNAS, FGF19, PTK2, FGFR1(10.00%); in brain metastases tissues were TP53(42.86%), PIK3CA(28.57%), ZNF703, PREX2, JAK3, MYC, FGFR1(14.29%). Besides, levels of TMB were also different among primary tissue, liver, lung and brain metastases tissues. Higher TMB level were showed in liver and brain metastases tissue than primary and lung metastases tissues (p&amp;lt;0.001). Conclusions: Different metastases of breast cancer were different in gene mutation landscape, this imply these patients should be managed differently. Citation Format: Yanman Fang, Hushan Zhang, Jiali Zou, Fulin Zhou, Xuejie Tang, Yi Yang, Li Luo, Yu Ding. Gene profile analysis of breast cancer lung metastasis, liver metastasis, brain metastasis and primary tumor [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5983.

Abstract 26: Influence of geographic/ancestral origin on glioma incidence and outcomes in US Hispanics

Abstract PURPOSE: Glioma is the most common type of primary malignant brain tumor, and incidence is 25% lower in U.S. Hispanics than White non-Hispanics. Though often pooled in epidemiological analyses, the U.S. Hispanic population is very diverse, and registry-based analyses may mask incidence and survival differences associated with geographic/ancestral origins. METHODS: County-level glioma incidence data in U.S. Hispanics from 2000-2017 were retrieved from the Central Brain Tumor Registry of the United States, and survival data from the CDC’s National Program of Cancer Registries was obtained for 2001-2017. American Community Survey data were used to determine county-level proportion of the Hispanic population of Mexican/Central-American origin and of Caribbean origin (Puerto Rican, Cuban, Dominican). State-level estimates of European and African admixture in Hispanics were obtained from published 23andMe data. Incidence rate ratios (IRRs) quantified the glioma incidence differences across groups. County-level incidence was modeled using linear regression to control for potential confounding factors (e.g., county size, median socioeconomic status), and Cox proportional hazards models were used to assess associations between individual-level survival, county-level origin percentages, and state-level admixture. RESULTS: Compared to Hispanics living in predominantly Caribbean-origin counties, Hispanics from predominantly Mexican/Central-American-origin counties had lower age-adjusted risks of glioma (IRR=0.83; P&amp;lt;0.0001), glioblastoma (IRR=0.86; P&amp;lt;0.0001), diffuse/anaplastic astrocytoma (IRR=0.78; P&amp;lt;0.0001), oligodendroglioma (IRR=0.82; P&amp;lt;0.0001), ependymoma (IRR=0.88; P=0.012), and pilocytic astrocytoma (IRR=0.76; P&amp;lt;0.0001). Associations were consistent in children and adults, and using more granular geographic regions. Despite having lower glioma incidence, Hispanic glioblastoma patients from predominantly Mexican/Central-American-origin counties had poorer survival than Hispanics living in predominantly Caribbean-origin counties. Incidence and survival differences could be partially explained by state-level estimates of European and African admixture in Hispanics, with European admixture associated with higher incidence and African admixture associated with improved survival. CONCLUSIONS: Glioma incidence and outcomes differ in association with the geographic origins of the Hispanic community, with residents of predominantly Mexican/Central-American-origin counties at significantly reduced risk and those of Caribbean-origin counties at comparatively greater risk. Although typically classified as a single ethnic group in registry data, appreciating the cultural, socioeconomic, and genetic diversity of U.S. Hispanics will help advance cancer disparities research. Citation Format: Kyle M. Walsh, Corey Neff, Melissa L. Bondy, Carol Kruchko, Jason T. Huse, Christopher I. Amos, Jill S. Barnholtz-Sloan, Quinn T. Ostrom. Influence of geographic/ancestral origin on glioma incidence and outcomes in US Hispanics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 26.

Abstract 770: Multiregional imaging and genomic analysis in high-grade IDH-mutant astrocytoma identifies intratumor subclones with aggressive features

Abstract High-grade gliomas represent the most common type of primary adult malignant brain tumor historically diagnosed and graded from histologic criteria alone. Gliomas harboring isocitrate dehydrogenase (IDH) 1 or 2 mutations, which are present in more than 80% of World Health Organization (WHO) grade 2 or 3 tumors, portend a favorable prognosis as compared to IDH-wildtype tumors. However, recent molecular profiling has identified the presence of additional alterations in these lower grade tumors that transform the biology toward an aggressive high-grade phenotype clinically that are independent of histologic grading and confer a worse clinical prognosis. Of these, the loss of CDKN2A/B is the strongest implicated alteration and is sufficient to define a grade 4 tumor in IDH-mutant astrocytomas regardless of histologic appearance. However, there remain no effective therapies targeted at molecular subgroups in aggressive gliomas to date. Here we have profiled the intratumoral heterogeneity via whole exome and RNA sequencing and copy number alteration analysis within a cohort of de-identified IDH-mutant astrocytoma patient tumors collected through the Mayo Clinic and Barrow Neurological Institute (BNI) biorepositories to determine the spatial evolution of aggressive molecular features. Within this cohort we have collected 42 image-localized intratumoral biopsy specimens representing 13 IDH-mutant patient tumors. We identified 11 biopsies from 3 tumors that harbored loss of CDKN2A/B which included 1 tumor that displayed intratumoral heterogeneity of CDKN2A/B expression across subclones. We performed an unsupervised cluster analysis to profile the transcriptional signatures in IDH-mutant astrocytomas across both differential CDKN2A/B status and tumor geographic location comparing contrast enhancing versus non-enhancing regions, and we analyzed the MRI features in IDH-mutant tumors across CDKN2A/B status. We found that tumors with CDKN2A/B alteration demonstrated lower signal intensity on delayed post-contrast T2*W images, which could represent lower pre-contrast T2*, unique contrast agent kinetics, or dissimilar diffusion properties related to increased cellular density and/or greater heterogeneity of cell size. Interestingly, we uncovered spatial heterogeneity of additional molecular drivers of high-grade glioma including KRAS, MYCN, and PDGFRA. While EGFR alterations are rarely reported in IDH-mutant tumors, we identified one treatment naïve patient tumor that harbored EGFR alteration only in the contrast non-enhancing region. Based on this work we propose there is early spatial evolution of aggressive tumor subclones prior to primary tumor diagnosis, and multiregional molecular profiling may better inform tumor biology and predict therapeutic strategies in IDH-mutant astrocytomas. Citation Format: Shannon P. Fortin Ensign, Fulvio D'Angelo, Francesca P. Caruso, Christopher Sereduk, Kyle Singleton, Jennifer Eschbacher, Gina Mazza, Maciej Mrugala, Alyx Porter, Bernard Bendok, Richard Zimmerman, Kris Smith, Peter Nakaji, Kamala Swanson, Junwen Wang, Nathenael Semmineh, Christopher Quarles, Anna Lasorella, Kristin Swanson, Michele Ceccarelli, Antonio Iavarone, Leland Hu, Nhan L. Tran. Multiregional imaging and genomic analysis in high-grade IDH-mutant astrocytoma identifies intratumor subclones with aggressive features [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 770.

Abstract 5329: Pt(IV)-M13, a blood-brain-barrier penetrant macrocyclic peptide-platinum(IV) conjugate, leads to enhanced drug uptake and efficacy in murine glioblastoma models

Abstract INTRODUCTION. Glioblastoma is the most aggressive primary malignant brain tumor and has a median survival of approximately 15 months. Treatment is hindered by the blood-brain barrier (BBB) that restricts entry of the vast majority of cancer therapeutics to the brain. We have designed a peptide-drug conjugate, (Pt(IV)-M13), comprised of M13, a perfluoroaryl-stapled cell-penetrating peptide, covalently linked to Pt(IV) via an amide bond. Upon intracellular reduction the Pt(IV) prodrug releases active cisplatin eliciting DNA damage and cell death. To investigate the properties of Pt(IV)-M13 in detail, we determined in vitro cell killing, platinum brain accumulation, biodistribution, and efficacy in mouse models. METHODS. Pt(IV)-M13 tumor cytotoxicity was assessed in vitro using standard approaches. In vivo platinum levels were measured in the brain by ICP-MS, and BBB penetration assessed through a multicellular 3D in vitro BBB model, and biodistribution to peripheral organs was determined. Toxicity from bi-weekly injections of the Pt(IV)-M13 platinum conjugate was monitored from 5mg/kg to 30mg/kg. Animal survival was determined in a glioblastoma xenograft model. DNA damage was quantified by γH2AX staining. RESULTS. Pt(IV)-M13 possesses in vitro tumor cell killing effects similar to cisplatin and Pt(IV) alone. Pt(IV)-M13 showed increased BBB penetration when compared to cisplatin in our in vitro BBB spheroid model (&amp;gt;20-fold), in brain tissue (7-fold) and glioblastoma tumor-bearing mice (8-fold). Five hours after intravenous injection, biodistribution of platinum delivered by Pt(IV)-M13 was higher than cisplatin in the spleen, but lower in the heart. Bi-weekly injections of Pt(IV)-M13 were well-tolerated in nude mice to at least 15mg/kg, three times the reported cisplatin MTD. Finally, Pt(IV)-M13 treatment increased survival in a murine glioblastoma xenograft model (median survival= 33 days for Pt(IV)-M13 vs. 22.5 days for cisplatin and 24 days Pt(IV) alone. Elevated γH2AX foci were observed in Pt(IV)-M13 treated mice providing direct support for the improved delivery of cisplatin to brain tumors using this approach. CONCLUSION. Pt(IV)-M13 leads to enhanced BBB penetration and cisplatin accumulation in glioblastoma models, with consequent animal survival benefit. Overall, our data support further investigation and development of Pt(IV)-M13 for glioblastoma treatment. Future studies will include additional glioblastoma models, and further optimization of this approach. Citation Format: Jorge-Luis Jimenez-Macias, Yen-Chun Lee, Tomer Finkelberg, Gilles Berger, Michal Nowicki, Choi-Fong Cho, Bogdan Fedeles, Andrei Loas, Brad Penetelute, Sean E. Lawler. Pt(IV)-M13, a blood-brain-barrier penetrant macrocyclic peptide-platinum(IV) conjugate, leads to enhanced drug uptake and efficacy in murine glioblastoma models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5329.

Abstract 3465: Efficacy of concomitant application of Tumor Treating Fields (TTFields), temozolomide and lomustine in glioblastoma cancer cells in vitro

Abstract Background: Glioblastoma (GBM) is the most common primary malignant brain tumor in adults. Newly diagnosed GBM (ndGBM) is treated with adjuvant temozolomide (TMZ), however, more than 50% of patients do not respond to therapy. Tumor Treating Fields (TTFields) therapy concomitant with maintenance TMZ was approved by the FDA in 2015 and represents a major advance in treatment of patients with ndGBM. Recently, lomustine (CCNU) together with TMZ demonstrated improved overall and progression-free survival following resection and radiochemotherapy in patients with ndGBM. Accordingly, the current study aimed to examine the effect of concomitant application of TTFields with TMZ and CCNU in GBM cells. Methods: GBM cells (U-87 MG and LN229) were treated for 72 h with TTFields at a frequency of 200 kHz and an intensity of 1 V/cm RMS using the inovitro system. Efficacy of combining TTFields with TMZ and/or CCNU was tested by measuring cell survival, clonogenic potential, overall effect (multiplicity product of cytotoxicity and clonogenicity), and apoptosis induction. Results: Combination of TMZ and TTFields resulted in increased cytotoxicity compared with each treatment alone. The cytotoxic effect resulting from co-application of CCNU with TTFields suggested a synergistic interaction between the two modalities. The clonogenic, overall, and apoptotic effects of TMZ and CCNU were also enhanced following co-treatment with TTFields. The triple combination of TTFields with both TMZ and CCNU was more efficacious than TMZ/CCNU or TTFields separately. Conclusions: The current research suggests potential benefits for the treatment of GBM using concomitant application of TTFields with TMZ/CCNU. Citation Format: Hila Fishman, Roni Monin, Eyal Dor-On, Adi Haber, Moshe Giladi, Uri Weinberg, Yoram Palti. Efficacy of concomitant application of Tumor Treating Fields (TTFields), temozolomide and lomustine in glioblastoma cancer cells in vitro [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3465.

#73 Single Cell RNA Sequencing of Zika Infection in Neural Tissues Reveals an Astrocyte-Driven Innate Immune Response Governed by Interferon Beta.

Abstract Background Congenital infection with Zika virus (ZIKV) has been linked to severe of fetal brain disruption and other developmental anomalies referred to collectively as congenital Zika syndrome (CZS). Long-term follow up of ZIKV-exposed infants has shown that even those without overt findings of CZS can have significant cognitive and developmental difficulties. However, the pathogenesis of ZIKV-related microcephaly and cognitive dysfunction remain poorly understood. To begin answering these questions, we sought to define the consequences of ZIKV infection on specific cell types in the developing brain. Method We have used primary human fetal brain cultures and human induced pluripotent stem cell-derived neural progenitor cell lines to probe innate immune signaling to ZIVK infection in vitro. We used single-cell RNA sequencing (10x Genomics) to identify the cell type-specific transcriptional changes induced by ZIKV infection. Results We found that neural progenitor cells (NPCs) support higher ZIKV RNA copy number than do differentiated neural cells. Moreover, while IFN-β is the predominant cytokine released by differentiated cells in response to ZIKV infection, NPCs do not produce IFN-β. By additionally characterizing the transcriptional response to IFN-β treatment, we have been able to distinguish virus-stimulated genes (VSGs) from interferon-stimulated genes (ISGs) across cell types. We find extensive heterogeneity in cell type-specific VSG and ISG profiles. Astrocytes undergo the largest transcriptional changes in response to infection and are the main producers of IFN-β. Our findings also identify genes whose expression is altered during viral infection specifically in NPCs and neurons, providing candidate pathways that may underlie neural dysfunction. Conclusion Across cell types, the response to Zika infection closely matches the response to IFN-β, arguing that astrocyte-derived IFN-β is the main driver of the innate immune response to ZIKV in the developing brain. Our findings provide insight into the pathogenesis of CZS and offer novel targets for improving developmental outcomes after fetal ZIKV infection.