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Papers by Marissa W. Madden

Research paper thumbnail of Ex vivo culture of circulating breast tumor cells for individualized testing of drug susceptibility

Research paper thumbnail of MAPK7 regulates EMT Features and Modulates the Generation of CTCs

Molecular cancer research : MCR, Jan 12, 2015

Epithelial-to-Mesenchymal Transition (EMT) has been implicated in models of tumor cell migration,... more Epithelial-to-Mesenchymal Transition (EMT) has been implicated in models of tumor cell migration, invasion and metastasis. In a search for candidate therapeutic targets to reverse this process, non-tumorigenic MCF10A breast epithelial cells were infected with an arrayed lentiviral kinome shRNA library and screened for either suppression or enhancement of a 26-gene EMT RNA signature. No individual kinase gene knockdown was sufficient to induce EMT. In contrast, grouped epithelial markers were induced by knockdown of multiple kinases, including mitogen activated protein kinase 7 (MAPK7). In breast cancer cells, suppression of MAPK7 increased E-cadherin (CDH1) expression and inhibited cell migration. In an orthotopic mouse model, MAPK7 suppression reduced the generation of circulating tumor cells (CTCs) and the appearance of lung metastases. Together, these observations raise the possibility that targeting kinases that maintain mesenchymal cell properties in cancer cells, such as MAPK7...

Research paper thumbnail of Brain tumor cells in circulation are enriched for mesenchymal gene expression

Cancer discovery, 2014

Glioblastoma (GBM) is a highly aggressive brain cancer characterized by local invasion and angiog... more Glioblastoma (GBM) is a highly aggressive brain cancer characterized by local invasion and angiogenic recruitment, yet metastatic dissemination is extremely rare. Here, we adapted a microfluidic device to deplete hematopoietic cells from blood specimens of patients with GBM, uncovering evidence of circulating brain tumor cells (CTC). Staining and scoring criteria for GBM CTCs were first established using orthotopic patient-derived xenografts (PDX), and then applied clinically: CTCs were identified in at least one blood specimen from 13 of 33 patients (39%; 26 of 87 samples). Single GBM CTCs isolated from both patients and mouse PDX models demonstrated enrichment for mesenchymal over neural differentiation markers compared with primary GBMs. Within primary GBMs, RNA in situ hybridization identified a subpopulation of highly migratory mesenchymal tumor cells, and in a rare patient with disseminated GBM, systemic lesions were exclusively mesenchymal. Thus, a mesenchymal subset of GBM c...

Research paper thumbnail of American Brain Tumor Association Young Investigator Award 198 Circulating Tumor Cells in Patients With Glioblastoma

Neurosurgery, 2014

ABSTRACT Glioblastoma (GBM) is characterized by necrosis, angiogenesis, and inevitable recurrence... more ABSTRACT Glioblastoma (GBM) is characterized by necrosis, angiogenesis, and inevitable recurrence. Despite its aggressiveness, GBM rarely forms extracranial metastases, suggesting impediments in vascular invasion, survival in circulation or implantation. Advances in microfluidics have successfully identified circulating tumor cells (CTCs) in lung, prostate, and breast cancer patients. Using a novel CTC microfluidic device, we hypothesize that GBM patients have CTCs, which can be captured, quantified, and analyzed for molecular and genetic markers.

Research paper thumbnail of Cancer therapy. Ex vivo culture of circulating breast tumor cells for individualized testing of drug susceptibility

Science (New York, N.Y.), Jan 11, 2014

Circulating tumor cells (CTCs) are present at low concentrations in the peripheral blood of patie... more Circulating tumor cells (CTCs) are present at low concentrations in the peripheral blood of patients with solid tumors. It has been proposed that the isolation, ex vivo culture, and characterization of CTCs may provide an opportunity to noninvasively monitor the changing patterns of drug susceptibility in individual patients as their tumors acquire new mutations. In a proof-of-concept study, we established CTC cultures from six patients with estrogen receptor-positive breast cancer. Three of five CTC lines tested were tumorigenic in mice. Genome sequencing of the CTC lines revealed preexisting mutations in the PIK3CA gene and newly acquired mutations in the estrogen receptor gene (ESR1), PIK3CA gene, and fibroblast growth factor receptor gene (FGFR2), among others. Drug sensitivity testing of CTC lines with multiple mutations revealed potential new therapeutic targets. With optimization of CTC culture conditions, this strategy may help identify the best therapies for individual canc...

Research paper thumbnail of Clusters of circulating tumor cells traverse capillary-sized vessels

Multicellular aggregates of circulating tumor cells (CTC clusters) are potent initiators of dista... more Multicellular aggregates of circulating tumor cells (CTC clusters)
are potent initiators of distant organ metastasis. However, it is
currently assumed that CTC clusters are too large to pass through
narrow vessels to reach these organs. Here, we present evidence
that challenges this assumption through the use of microfluidic
devices designed to mimic human capillary constrictions and CTC
clusters obtained from patient and cancer cell origins. Over 90% of
clusters containing up to 20 cells successfully traversed 5- to 10-μm
constrictions even in whole blood. Clusters rapidly and reversibly
reorganized into single-file chain-like geometries that substantially
reduced their hydrodynamic resistances. Xenotransplantation of human
CTC clusters into zebrafish showed similar reorganization and
transit through capillary-sized vessels in vivo. Preliminary experiments
demonstrated that clusters could be disrupted during transit using
drugs that affected cellular interaction energies. These findings
suggest that CTC clusters may contribute a greater role to tumor
dissemination than previously believed and may point to strategies
for combating CTC cluster-initiated metastasis.

Research paper thumbnail of Ex vivo culture of circulating breast tumor cells for individualized testing of drug susceptibility

Research paper thumbnail of MAPK7 regulates EMT Features and Modulates the Generation of CTCs

Molecular cancer research : MCR, Jan 12, 2015

Epithelial-to-Mesenchymal Transition (EMT) has been implicated in models of tumor cell migration,... more Epithelial-to-Mesenchymal Transition (EMT) has been implicated in models of tumor cell migration, invasion and metastasis. In a search for candidate therapeutic targets to reverse this process, non-tumorigenic MCF10A breast epithelial cells were infected with an arrayed lentiviral kinome shRNA library and screened for either suppression or enhancement of a 26-gene EMT RNA signature. No individual kinase gene knockdown was sufficient to induce EMT. In contrast, grouped epithelial markers were induced by knockdown of multiple kinases, including mitogen activated protein kinase 7 (MAPK7). In breast cancer cells, suppression of MAPK7 increased E-cadherin (CDH1) expression and inhibited cell migration. In an orthotopic mouse model, MAPK7 suppression reduced the generation of circulating tumor cells (CTCs) and the appearance of lung metastases. Together, these observations raise the possibility that targeting kinases that maintain mesenchymal cell properties in cancer cells, such as MAPK7...

Research paper thumbnail of Brain tumor cells in circulation are enriched for mesenchymal gene expression

Cancer discovery, 2014

Glioblastoma (GBM) is a highly aggressive brain cancer characterized by local invasion and angiog... more Glioblastoma (GBM) is a highly aggressive brain cancer characterized by local invasion and angiogenic recruitment, yet metastatic dissemination is extremely rare. Here, we adapted a microfluidic device to deplete hematopoietic cells from blood specimens of patients with GBM, uncovering evidence of circulating brain tumor cells (CTC). Staining and scoring criteria for GBM CTCs were first established using orthotopic patient-derived xenografts (PDX), and then applied clinically: CTCs were identified in at least one blood specimen from 13 of 33 patients (39%; 26 of 87 samples). Single GBM CTCs isolated from both patients and mouse PDX models demonstrated enrichment for mesenchymal over neural differentiation markers compared with primary GBMs. Within primary GBMs, RNA in situ hybridization identified a subpopulation of highly migratory mesenchymal tumor cells, and in a rare patient with disseminated GBM, systemic lesions were exclusively mesenchymal. Thus, a mesenchymal subset of GBM c...

Research paper thumbnail of American Brain Tumor Association Young Investigator Award 198 Circulating Tumor Cells in Patients With Glioblastoma

Neurosurgery, 2014

ABSTRACT Glioblastoma (GBM) is characterized by necrosis, angiogenesis, and inevitable recurrence... more ABSTRACT Glioblastoma (GBM) is characterized by necrosis, angiogenesis, and inevitable recurrence. Despite its aggressiveness, GBM rarely forms extracranial metastases, suggesting impediments in vascular invasion, survival in circulation or implantation. Advances in microfluidics have successfully identified circulating tumor cells (CTCs) in lung, prostate, and breast cancer patients. Using a novel CTC microfluidic device, we hypothesize that GBM patients have CTCs, which can be captured, quantified, and analyzed for molecular and genetic markers.

Research paper thumbnail of Cancer therapy. Ex vivo culture of circulating breast tumor cells for individualized testing of drug susceptibility

Science (New York, N.Y.), Jan 11, 2014

Circulating tumor cells (CTCs) are present at low concentrations in the peripheral blood of patie... more Circulating tumor cells (CTCs) are present at low concentrations in the peripheral blood of patients with solid tumors. It has been proposed that the isolation, ex vivo culture, and characterization of CTCs may provide an opportunity to noninvasively monitor the changing patterns of drug susceptibility in individual patients as their tumors acquire new mutations. In a proof-of-concept study, we established CTC cultures from six patients with estrogen receptor-positive breast cancer. Three of five CTC lines tested were tumorigenic in mice. Genome sequencing of the CTC lines revealed preexisting mutations in the PIK3CA gene and newly acquired mutations in the estrogen receptor gene (ESR1), PIK3CA gene, and fibroblast growth factor receptor gene (FGFR2), among others. Drug sensitivity testing of CTC lines with multiple mutations revealed potential new therapeutic targets. With optimization of CTC culture conditions, this strategy may help identify the best therapies for individual canc...

Research paper thumbnail of Clusters of circulating tumor cells traverse capillary-sized vessels

Multicellular aggregates of circulating tumor cells (CTC clusters) are potent initiators of dista... more Multicellular aggregates of circulating tumor cells (CTC clusters)
are potent initiators of distant organ metastasis. However, it is
currently assumed that CTC clusters are too large to pass through
narrow vessels to reach these organs. Here, we present evidence
that challenges this assumption through the use of microfluidic
devices designed to mimic human capillary constrictions and CTC
clusters obtained from patient and cancer cell origins. Over 90% of
clusters containing up to 20 cells successfully traversed 5- to 10-μm
constrictions even in whole blood. Clusters rapidly and reversibly
reorganized into single-file chain-like geometries that substantially
reduced their hydrodynamic resistances. Xenotransplantation of human
CTC clusters into zebrafish showed similar reorganization and
transit through capillary-sized vessels in vivo. Preliminary experiments
demonstrated that clusters could be disrupted during transit using
drugs that affected cellular interaction energies. These findings
suggest that CTC clusters may contribute a greater role to tumor
dissemination than previously believed and may point to strategies
for combating CTC cluster-initiated metastasis.