Noninvasively Delivering Therapeutic Monoclonal Antibodies into the Brain–Opportunity in Primary Brain Tumor/ Breast Cancer Brain Metastasis Treatment

ABSTRACTBreast cancer brain metastasis remains largely incurable. While several mouse models have been developed to investigate the genes and mechanisms regulating breast cancer brain metastasis, these models often lack clinical relevance since they require the use of immune-compromised mice and/or are poorly metastatic to brain from the mammary gland. We describe the development and characterisation of an aggressive brain metastatic variant of the 4T1 syngeneic model (4T1Br4) that spontaneously metastasises to multiple organs, but is selectively more metastatic to the brain from the mammary gland than parental 4T1 tumours. By immunohistochemistry, 4T1Br4 tumours and brain metastases display a triple negative phenotype, consistent with the high propensity of this breast cancer subtype to spread to brain. In vitro assays indicate that 4T1Br4 cells have an enhanced ability to adhere to or migrate across a brain-derived endothelial monolayer and greater invasive response to brain-derived soluble factors compared to 4T1 cells. These properties are likely to contribute to the brain-selectivity of 4T1Br4 tumours. Expression profiling and gene set enrichment analyses demonstrate the clinical relevance of the 4T1Br4 model at the transcriptomic level. Pathway analyses implicate tumour-intrinsic immune regulation and vascular interactions in successful brain colonisation, revealing potential therapeutic targets. Evaluation of two histone deacetylase inhibitors, SB939 and 1179.4b, shows partial efficacy against 4T1Br4 metastasis to brain and other sites in vivo and potent radio-sensitising properties in vitro. The 4T1Br4 model provides a clinically relevant tool for mechanistic studies and to evaluate novel therapies against brain metastasis.SUMMARY STATEMENTWe introduce a new syngeneic mouse model of spontaneous breast cancer brain metastasis, demonstrate its phenotypic, functional and transcriptomic relevance to human TNBC brain metastasis and test novel therapies.

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Glucose Transporter 3 Is Essential for the Survival of Breast Cancer Cells in the Brain

Cells ◽

10.3390/cells8121568 ◽

2019 ◽

Vol 8 (12) ◽

pp. 1568 ◽

Cited By ~ 1

Author(s):

Min-Hsun Kuo ◽

Wen-Wei Chang ◽

Bi-Wen Yeh ◽

Yeh-Shiu Chu ◽

Yueh-Chun Lee ◽

...

Keyword(s):

Breast Cancer ◽

Glucose Metabolism ◽

Brain Metastasis ◽

Cancer Cells ◽

Breast Cancer Cells ◽

Glucose Transporter ◽

Metastatic Breast ◽

Breast Cancer Brain Metastasis ◽

The Brain

Breast cancer brain metastasis commonly occurs in one-fourth of breast cancer patients and is associated with poor prognosis. Abnormal glucose metabolism is found to promote cancer metastasis. Moreover, the tumor microenvironment is crucial and plays an active role in the metabolic adaptations and survival of cancer cells. Glucose transporters are overexpressed in cancer cells to increase glucose uptake. The glucose transporter 3 (GLUT3) is a high-affinity glucose transporter that is highly expressed in mammalian neurons. GLUT3 is also overexpressed in several malignant brain tumors. However, the role of GLUT3 in breast cancer brain metastasis remains unknown. The results of the present study demonstrated that GLUT3 is highly overexpressed in brain metastatic breast cancers and mediates glucose metabolic reprogramming. Furthermore, knockdown of cAMP-response element binding protein (CREB) could directly regulate GLUT3 expression in brain metastatic breast cancer cells. Notably, we verified and provided a novel role of GLUT3 in mediating glucose metabolism and assisting breast cancer cells to survive in the brain to promote brain metastasis.

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TAMI-04. OLFACTORY RECEPTOR 5B21 DRIVES BREAST CANCER BRAIN METASTASIS THROUGH STAT3/NFΚB/CEBPΒ PATHWAY

Neuro-Oncology ◽

10.1093/neuonc/noab196.788 ◽

2021 ◽

Vol 23 (Supplement_6) ◽

pp. vi198-vi199

Author(s):

Mao Li ◽

Markus Schweiger ◽

Daniel Ryan ◽

Ichiro Nakano ◽

Litia Carvalho ◽

...

Keyword(s):

Breast Cancer ◽

Brain Metastasis ◽

Cancer Metastasis ◽

Metastatic Breast ◽

Mesenchymal Transition ◽

Breast Cancer Brain Metastasis ◽

Metastatic Sites ◽

The Brain

Abstract Olfactory receptors (ORs), responsible for the sense of smell, play an essential role in various physiological processes outside the nasal epithelium, including cancer. In breast cancer, however, the expression and function of ORs remain understudied. We established a breast cancer metastasis model by intracardiac injection of MDA-MB-231 (231P) in immunocompromised mice and produced a series of derivative cell lines from developed metastatic sites, including the brain-seeking clone (231Br). We examined the significance of ORs transcript abundance in primary and metastatic breast cancer to different tissues, including the brain, bone, and lung. While 20 OR transcripts were differentially expressed in distant metastases, OR5B21 displayed high expression in all three metastatic sites with respect to the primary tumor, especially in brain metastasis with 13 fold higher than the primary site. Metastatic clones showed distinguishing higher invasion biological characteristics compared to parental cells in vivo and in vitro. Knockdown of OR5B21 significantly decreased the invasion and migration of MDA-MB-231 Brain-seeking metastatic cell as well as metastasis to different organs, including the brain, while overexpression of OR5B21 had the opposite effect. Mechanistically, OR5B21 expression was associated with epithelial to mesenchymal transition through the STAT3/NFkB/CEBPβ signaling pathway. We propose OR5B21 (and potentially other ORs) as a novel oncogene contributing to breast cancer brain metastasis and a potential target for adjuvant therapy.

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Evolution of the Tumor Microenvironment toward Immune-Suppressive Seclusion during Brain Metastasis of Breast Cancer: Implications for Targeted Therapy

Cancers ◽

10.3390/cancers13194895 ◽

2021 ◽

Vol 13 (19) ◽

pp. 4895

Author(s):

Myung-Giun Noh ◽

Sung Sun Kim ◽

Yeong Jin Kim ◽

Tae-Young Jung ◽

Shin Jung ◽

...

Keyword(s):

Breast Cancer ◽

Tumor Microenvironment ◽

Brain Metastasis ◽

Immune Cell ◽

Metastatic Cancer ◽

Emerging Therapies ◽

M1 Macrophage ◽

Breast Cancer Brain Metastasis ◽

New Treatments ◽

The Brain

Breast cancer (BC) is the second most common solid malignant tumor that metastasizes to the brain. Despite emerging therapies such as immunotherapy, whether the tumor microenvironment (TME) in breast cancer brain metastasis (BCBM) has potential as a target of new treatments is unclear. Expression profiling of 770 genes in 12 pairs of primary BC and matched brain metastasis (BM) samples was performed using the NanoString nCounter PanCancer IO360TM Panel. Immune cell profiles were validated by immunohistochemistry (IHC) in samples from 50 patients with BCBM. Pathway analysis revealed that immune-related pathways were downregulated. Immune cell profiling showed that CD8+ T cells and M1 macrophages were significantly decreased, and M2 macrophages were significantly increased, in BM compared to primary BC samples (p = 0.001, p = 0.021 and p = 0.007, respectively). CCL19 and CCL21, the top differentially expressed genes, were decreased significantly in BM compared to primary BC (p < 0.001, both). IHC showed that the CD8+ count was significantly lower (p = 0.027), and the CD163+ and CD206+ counts were higher, in BM than primary BC (p < 0.001, both). A low CD8+ T cell count, low CD86+ M1 macrophage count, and high M2/M1 macrophage ratio were related to unfavorable clinical outcomes. BC exhibits an immunosuppressive characteristic after metastasis to the brain. These findings will facilitate establishment of a treatment strategy for BCBM based on the TME of metastatic cancer.

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EXTH-02. TUMOR-HOMING INDUCED NEURAL STEM CELL THERAPY INHIBITS THE PROGRESSION OF BREAST CANCER BRAIN METASTASIS AND LEPTOMENINGEAL CARCINOMATOSIS

Neuro-Oncology ◽

10.1093/neuonc/noaa215.356 ◽

2020 ◽

Vol 22 (Supplement_2) ◽

pp. ii86-ii87

Author(s):

Wulin Jiang ◽

Alain Valdivia ◽

Alison Mercer-Smith ◽

Carey Anders ◽

Shawn Hingtgen

Keyword(s):

Breast Cancer ◽

Brain Metastasis ◽

Median Survival ◽

Nude Mice ◽

Limit Of Detection ◽

Human Fibroblasts ◽

Leptomeningeal Carcinomatosis ◽

Breast Cancer Brain Metastasis ◽

Tumor Homing ◽

The Brain

Abstract INTRODUCTION Breast cancer brain metastasis, including leptomeningeal carcinomatosis (LC), remains one of the most lethal CNS diseases. New therapies are urgently needed to treat this highly aggressive disease. Here we used models of both breast cancer brain parenchymal metastasis and leptomeningeal metastasis to investigate the efficacy of engineered tumor-homing neural stem cells (NSCs) therapy. METHODS Personalized NSCs were created using Sox2 overexpression to transdifferentiate human fibroblasts into induced NSCs (iNSCs), followed by genetic engineering to enable iNSCs to secrete cytotoxic TRAIL (iNSC-TRAIL). For the parenchymal metastasis study, iNSC-TRAIL therapy was infused intracerebroventricularly (ICV) into Nude mice bearing established intracranial MDA-MB-231-Br human breast cancer cells expressing fluorescent and bioluminescent reporters. For LC studies, we established the disease model by inoculating Nude mice with MDA-MB-231-Br tumor cells via intracisternal infusion. iNSC-TRAIL therapy was evaluated by infusing therapy ICV either 1 week prior to or 3 days after tumor inoculation to mirror prophylactic or established tumor treatment, respectively. Tumor progression in the brain and spine was monitored by serial bioluminescence imaging (BLI), and survival was analyzed. RESULTS Serial BLI showed ICV-infused iNSC-TRAIL reduced parenchymal tumor volumes by 72% 3 weeks post-ICV infusion, and extended median survival from 37 to 52 days. Testing iNSC-TRAIL therapy against established LC tumors, serial BLI showed ICV iNSC-TRAIL therapy reduced established tumors 196-fold in the brain and 500-fold in the spine within 2 weeks post-infusion, while extending median survival from 25 to 47 days. In the prophylactic LC model, iNSC-TRAIL therapy markedly delayed tumor development with tumors in the brain remaining > 1000-fold smaller than control, and tumors in the spine below the limit of detection through 1 month post-treatment. The therapy also eliminated mortality through 50 days post-therapy. CONCLUSION These data suggest iNSC therapy could be a promising treatment option for breast cancer brain metastasis patients.

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Capturing Changes in the Brain Microenvironment during Initial Steps of Breast Cancer Brain Metastasis

American Journal Of Pathology ◽

10.2353/ajpath.2010.090838 ◽

2010 ◽

Vol 176 (6) ◽

pp. 2958-2971 ◽

Cited By ~ 163

Author(s):

Mihaela Lorger ◽

Brunhilde Felding-Habermann

Keyword(s):

Breast Cancer ◽

Brain Metastasis ◽

Breast Cancer Brain Metastasis ◽

The Brain

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Novel Breast Cancer Brain Metastasis Patient-Derived Orthotopic Xenograft Model for Preclinical Studies

Cancers ◽

10.3390/cancers12020444 ◽

2020 ◽

Vol 12 (2) ◽

pp. 444 ◽

Cited By ~ 4

Author(s):

Masanori Oshi ◽

Maiko Okano ◽

Aparna Maiti ◽

Omar M. Rashid ◽

Katsuharu Saito ◽

...

Keyword(s):

Breast Cancer ◽

Brain Metastasis ◽

Tumor Heterogeneity ◽

Xenograft Model ◽

Orthotopic Xenograft Model ◽

Patient Derived Xenograft ◽

Pdx Model ◽

Breast Cancer Brain Metastasis ◽

New Treatments ◽

The Brain

The vast majority of mortality in breast cancer results from distant metastasis. Brain metastases occur in as many as 30% of patients with advanced breast cancer, and the 1-year survival rate of these patients is around 20%. Pre-clinical animal models that reliably reflect the biology of breast cancer brain metastasis are needed to develop and test new treatments for this deadly condition. The patient-derived xenograft (PDX) model maintains many features of a donor tumor, such as intra-tumor heterogeneity, and permits the testing of individualized treatments. However, the establishment of orthotopic PDXs of brain metastasis is procedurally difficult. We have developed a method for generating such PDXs with high tumor engraftment and growth rates. Here, we describe this method and identify variables that affect its outcomes. We also compare the brain-orthotopic PDXs with ectopic PDXs grown in mammary pads of mice, and show that the responsiveness of PDXs to chemotherapeutic reagents can be dramatically affected by the site that they are in.

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Rab11b-mediated integrin recycling promotes brain metastatic adaptation and outgrowth

10.1101/666750 ◽

2019 ◽

Author(s):

Erin N. Howe ◽

Miranda D. Burnette ◽

Melanie E. Justice ◽

James W. Clancy ◽

Ian H. Guldner ◽

...

Keyword(s):

Breast Cancer ◽

Brain Metastasis ◽

Mevalonate Pathway ◽

Surface Expression ◽

Integrin Β1 ◽

Breast Cancer Brain Metastasis ◽

Cargo Proteins ◽

Breast Cancer Brain Metastases ◽

Surface Proteome ◽

The Brain

GRAPHICAL ABSTRACTSUMMARYBreast cancer brain metastases (BCBM) have a 5-20 year latency and account for up to 30% of mortality. Developing new therapeutics requires a molecular understanding of adaptation to the brain microenvironment. Here, we combined RNA-sequencing of BCBM development with a reverse genetic screen in Drosophila melanogaster and identified Rab11b, an endosomal recycling protein, as a mediator of metastatic adaptation. We show that disseminated cells up-regulate Rab11b early after arrival in the brain, allowing control of the cell surface proteome through recycling of proteins required for successful interaction with the microenvironment, including integrin β1. Rab11b-mediated control of integrin β1 surface expression allows ligation to the brain ECM, activating mechanotransduction signaling to allow survival and proliferation. We propose a model in which up-regulation of Rab11b allows disseminated cells to recycle needed proteins during metastatic adaptation, without strictly requiring transcription and translation, to allow for metastatic outgrowth.Manuscript SummaryRab11b up-regulation in the brain microenvironment promotes recycling of cargo proteins required for breast cancer brain metastasis, including increased surface expression of integrin β1, which allows brain extracellular matrix attachment and mechanotransduction. Inhibition of the mevalonate pathway with statins prevents geranylgeranylation of Rab11b, decreasing cargo recycling, and inhibiting brain metastasis.

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