ABSTRACT
The PI, Irida Kastrati, PhD, transferred from the University of Illinois at Chicago to a new institution, Loyola University Chicago (LUC).At LUC, the PI obtained a new IACUC approval and completed an HRPO review. An MTA was submitted and is pending approval for transferring PDXs from our collaborator Dr. Carol Sartorius at U. Colorado to our institution. Initiation of in vivo animal work is currently halted due to COVID-19 measures in Illinois.
ABSTRACT
This project centers on the NF1/neurofibromin tumor suppressor, which was best known as a GTPase Activating Protein (GAP)that repress Ras activity. The parent DoD award has successfully defined a new and GAP-independent activity that NF1 is also a transcriptional co-repressor for estrogen receptor (ER) in ER+ breast cancer. While the parent DoD award focused on endocrine therapy resistance caused by NF1 loss, in this Expansion Award, the focus instead is on metastasis, for which currently has no cure. An important feature of ER+ breast cancer metastasis is that greater than 70% of the metastasis is in the bone. We hypothesized that the transcriptional co-repressor role of NF1 is also responsible for driving bone metastasis in ER+ breast cancer. Therefore, the objective of this Expansion Award is to assess NF1s role in metastasis in order to establish a strategy to stop it. We have made progress in accomplish Task1/Aim 1 to fully define NF1-controlled genes that can impact bone metastasis. This was a key part of the data that was just published in the high impact journal Cancer Cell. This award has also supported the launching a Phase-II clinical trial to treat ER+ NF1-depleted breast cancer, and the awards of a SPORE and another DoD Level-2 project. However, in Aim 2 (Tasks 2 and 30) we are dependent on the use of animals to study how NF1-depleted cancer cells interact with the bone, but this line of study has been severely and negatively impacted by COVID-19. We discuss how we plan to overcome this problem in the future.
ABSTRACT
AIMS: There are approximately four thousand neuro-oncology procedures in the UK per annum. Many of these result in tissue and biofluid specimens that are surplus to diagnostic requirement and can be collected as standard of clinical care. However, developing technologies and treatments for precision medicine require access to a range of individualised biospecimens paired with deep clinical phenotyping data. Here, we present Brain Surgical Tissue for Advanced Tumour Models (BRAINSTAT) programme, an infrastructure that has been established between Queen Elizabeth Hospital, Birmingham and the University of Birmingham, to collect, structure and store these resources and also maximise their value for research over the long-term. Using this approach our aim is to provide high-quality, annotated resources to help develop novel treatments for patients with brain tumours. METHOD: BRAINSTAT infrastructure allows: Prospective consent Biospecimens, including tumour tissue (brain and other primary in the case of metastasis), cyst fluid, dura, skin, CSF, blood (matched "germ-line" and for circulating cell free tumour DNA analysis), urine and saliva can be collected. Consent for long term follow-up, is either via clinic or NHS digital. More limited consent for non-oncological neurosurgical cohorts (e.g. epilepsy or vascular) and healthy volunteers allow healthy access-tissue and biofluids to be collected. B. Rapid transfer of fresh surgical tissue samples: Strong collaborative links and close physical proximity between operating theatre and laboratory allows rapid transfer of biospecimens minimising transit time. C. Standardised annotation across disciplines The RedCAP database system allows granular control over data-access, and each specialist research team is provided access only to the sub-sections relevant to them. All users must have Good Clinical Practice certification and GDPR training, prior to access of the BRAINSTAT database. RESULTS: Between 25/11/2019-16/03/2020 and 27/07/2020-16/11/2020, 65 patients were consented for BRAINSTAT at the weekly neurosurgical oncology clinic. (Recruitment gaps due to the SARS-COVID 19 pandemic). Pathological diagnosis of surplus tissue collected included: 37 high grade glioma, 3 low grade glioma and 16 brain metastasis including: (6 lung, 6 breast, 2 colorectal, 1 oesophageal, 1 endometrial). Meningioma (5 WHO I;1 WHO III) 1 patient undergoing anterior temporal lobectomy for hippocampal sclerosis contributed access tissue from the lateral neocortex. 1 patient had a non-neoplastic, non-diagnostic sample. All patients had matched "germline" blood samples. Median time from resection to arrival in the laboratory was 10 minutes (range 4-31). Standardised operating protocols to optimise this have been developed. Glioblastoma and breast-brain metastasis tumourspheres and cerebral organoids are currently being validated. CONCLUSION: Despite the challenges of the pandemic we have established a viable tissue pipeline from neurosurgical operating theatre to our university laboratories. We are developing clinically annotated human brain tumour cell lines, stem cells and 3D organoid models, principally for commonly encountered brain tumours such as glioma and metastasis. The research sets the foundation for a multitude of downstream applications including:-Building more complex organoid cultures e.g. by including other cell types such as healthy brain cells and endothelial cells allowing future experiments to more accurately model tumour growth.-Developing high-throughput, patient-specific drug screens of novel drugs and drug combinations using these 3D tumour models aiming to more effectively treat tumour proliferation and spread. These patient avatars will help inform and test more "stratified" personal medical treatments and will provide opportunities to allow earlier intervention with the aim of improving survival, coupled with a better quality of life.