- Email: [email protected]
Identification of genes associated with breast cancer metastatic to bone marrow
S40
Multi-Disciplinary Session I
expression of MCM7 was down regulated by 50% and 20% at 17 and 24 weeks, respectively. CONCLUSION(S): Through molecular fingerprinting, we have confirmed a potential role for MCM7 in the development of prostate cancer, demonstrating over-expression at 12 weeks prior to the histologic appearance of tumor. Moreover, we have identified MCM7 as a transcriptional target of green tea, and a potential molecular mediator of its chemo-preventive effect.
A translational approach to modeling pneumonia dynamics Jonathan E McDunn PhD, Ashoka Polpitiya DSc, Gary Stormo PhD, Bijoy Ghosh PhD, J Perren Cobb MD, FACS Washington University School of Medicine, Saint Louis, MO INTRODUCTION: We have shown that circulating leukocyte microarray profiles distinguish between the host response to Grampositive and Gram-negative pneumonia in mice at 24 hours. We hypothesized that similar microarray profiles could model the dynamic human response to pneumonia. METHODS: Mice underwent intratracheal (i.t.) injections of P. aeruginosa, S. pneumoniae or normal saline. GeneChip analysis of circulating leukocyte RNA identified twenty-seven genes that distinguished between the two infections. Similarly, leukocyte microarray expression profiles were generated from mechanically ventilated patients every two days for three weeks. Patients developed nosocomial pneumonia within ten days of being ventilated. Homologs for 16 of the 27 genes identified in the murine study were represented on the human microarray. RESULTS: Single-value decomposition of the expression values of these 16 genes generated patient-specific 21-day trajectories describing the host response to the onset of and convalescence from pneumonia. These molecular trajectories have a ten-fold greater signal than trajectories created from 16 randomly selected genes. CONCLUSIONS: Here we present the culmination of a mouse pneumonia study, demonstrating that the model directly informs analysis of a corresponding clinical study. Furthermore, we demonstrate for the first time that disease trajectories derived from microarray expression profiles can be used to model the clinical course of acute disease. These data suggest that leukocyte microarray profiles can be used as a clinical tool to quantitatively define the dynamics of pathophysiologic states.
Identification of genes associated with breast cancer metastatic to bone marrow Tara L Huston MD, Baixin Zhu MD, Michael Osborne MD, FRCS, FACS, Rache Simmons MD, Thomas Fahey III MD, FACS New York Presbyterian Hospital/Weill Cornell Medical Center, New York, NY INTRODUCTION: Bone marrow (BM) micrometastases are a marker for early hematogenous cancer dissemination in breast carci-
J Am Coll Surg
noma. Numerous studies have established that documentation of BM micrometastases at the time of initial diagnosis is as important as lymph node metastases when determining prognosis. However, the bone marrow is rarely examined in patients with breast cancer. These studies were undertaken to determine whether molecular profiling could identify primary tumors with associated BM micrometastases. METHODS: Molecular profiling by oligonucleotide microarray analysis was performed on twenty-two invasive ductal carcinomas from patients without overt metastases, who underwent resection of their primary tumor. Specimens were divided based on results of lymph node sampling and bone marrow aspiration: 1. LN-, BM-; 2. LN⫹, BM-; 3. LN-, BM⫹; and 4. LN⫹, BM⫹. Hierarchical cluster analysis defined potential groupings and differences among samples that spread either lymphatically, hematogenously or both. RESULTS: Using cDNA microarray technology, a list of 74 differentially expressed genes was created (p⬍0.01). Cancers with BM micrometastases clustered as a single group, while those without BM micrometastases clustered separately. The profiles of tumors with BM micrometastases were characterized by downregulation of tumor suppressor genes rather than upregulation of growth related genes. Interestingly, the profile included genes involved in aspects of malignant transformation, including angiogenesis, basement membrane invasion and cell migration. CONCLUSIONS: Bone marrow positive breast cancers can be identified by molecular profiling of the primary tumor. Diagnosis of BMM by molecular profiling of the primary tumor may improve staging and thus adjuvant treatment of patients with breast cancer.
Leptin regulates gallbladder genes related to gallstone pathogenesis Shannon J Graewin MD, James Kiely MD, Deborah Swartz-Basile PhD, Carol Svatek BS, Hayder Al-Azzawi MD, Henry Pitt MD, FACS Medical College of Wisconsin, Milwaukee, WI INTRODUCTION: Cholesterol gallstone formation requires three factors: abnormal gallbladder motility, enhanced cholesterol crystallization, and supersaturated bile. We have reported that obese, leptindeficient and leptin-resistant mice have abnormal gallbladder emptying and altered cholesterol crystal formation. But the genetic factors related to gallbladder motility and crystal formation have not been elucidated. The aim of this study was to identify which gallbladder genes related to gallstone pathogenesis are regulated by leptin. METHODS: Twenty-four eight-week old leptin-deficient mice were fed a non-lithogenic diet for four weeks. 12 mice received daily IP saline injections while 12 received 5 g/g recombinant leptin. Gallbladder mRNA was isolated, pooled (4 saline, 4 leptin) and analyzed on murine microarray chips. RESULTS: The leptin-deficient mice receiving leptin had significant weight loss (34 vs 49g, p⬍0.001) and reduction in gallbladder volume (17 vs 42 l, p⬍0.001). They also had significant upregulation of the leptin receptor (p⬍0.01), JAK 1 (p⬍0.001), STAT 1 and
Multi-Disciplinary Session I
expression of MCM7 was down regulated by 50% and 20% at 17 and 24 weeks, respectively. CONCLUSION(S): Through molecular fingerprinting, we have confirmed a potential role for MCM7 in the development of prostate cancer, demonstrating over-expression at 12 weeks prior to the histologic appearance of tumor. Moreover, we have identified MCM7 as a transcriptional target of green tea, and a potential molecular mediator of its chemo-preventive effect.
A translational approach to modeling pneumonia dynamics Jonathan E McDunn PhD, Ashoka Polpitiya DSc, Gary Stormo PhD, Bijoy Ghosh PhD, J Perren Cobb MD, FACS Washington University School of Medicine, Saint Louis, MO INTRODUCTION: We have shown that circulating leukocyte microarray profiles distinguish between the host response to Grampositive and Gram-negative pneumonia in mice at 24 hours. We hypothesized that similar microarray profiles could model the dynamic human response to pneumonia. METHODS: Mice underwent intratracheal (i.t.) injections of P. aeruginosa, S. pneumoniae or normal saline. GeneChip analysis of circulating leukocyte RNA identified twenty-seven genes that distinguished between the two infections. Similarly, leukocyte microarray expression profiles were generated from mechanically ventilated patients every two days for three weeks. Patients developed nosocomial pneumonia within ten days of being ventilated. Homologs for 16 of the 27 genes identified in the murine study were represented on the human microarray. RESULTS: Single-value decomposition of the expression values of these 16 genes generated patient-specific 21-day trajectories describing the host response to the onset of and convalescence from pneumonia. These molecular trajectories have a ten-fold greater signal than trajectories created from 16 randomly selected genes. CONCLUSIONS: Here we present the culmination of a mouse pneumonia study, demonstrating that the model directly informs analysis of a corresponding clinical study. Furthermore, we demonstrate for the first time that disease trajectories derived from microarray expression profiles can be used to model the clinical course of acute disease. These data suggest that leukocyte microarray profiles can be used as a clinical tool to quantitatively define the dynamics of pathophysiologic states.
Identification of genes associated with breast cancer metastatic to bone marrow Tara L Huston MD, Baixin Zhu MD, Michael Osborne MD, FRCS, FACS, Rache Simmons MD, Thomas Fahey III MD, FACS New York Presbyterian Hospital/Weill Cornell Medical Center, New York, NY INTRODUCTION: Bone marrow (BM) micrometastases are a marker for early hematogenous cancer dissemination in breast carci-
J Am Coll Surg
noma. Numerous studies have established that documentation of BM micrometastases at the time of initial diagnosis is as important as lymph node metastases when determining prognosis. However, the bone marrow is rarely examined in patients with breast cancer. These studies were undertaken to determine whether molecular profiling could identify primary tumors with associated BM micrometastases. METHODS: Molecular profiling by oligonucleotide microarray analysis was performed on twenty-two invasive ductal carcinomas from patients without overt metastases, who underwent resection of their primary tumor. Specimens were divided based on results of lymph node sampling and bone marrow aspiration: 1. LN-, BM-; 2. LN⫹, BM-; 3. LN-, BM⫹; and 4. LN⫹, BM⫹. Hierarchical cluster analysis defined potential groupings and differences among samples that spread either lymphatically, hematogenously or both. RESULTS: Using cDNA microarray technology, a list of 74 differentially expressed genes was created (p⬍0.01). Cancers with BM micrometastases clustered as a single group, while those without BM micrometastases clustered separately. The profiles of tumors with BM micrometastases were characterized by downregulation of tumor suppressor genes rather than upregulation of growth related genes. Interestingly, the profile included genes involved in aspects of malignant transformation, including angiogenesis, basement membrane invasion and cell migration. CONCLUSIONS: Bone marrow positive breast cancers can be identified by molecular profiling of the primary tumor. Diagnosis of BMM by molecular profiling of the primary tumor may improve staging and thus adjuvant treatment of patients with breast cancer.
Leptin regulates gallbladder genes related to gallstone pathogenesis Shannon J Graewin MD, James Kiely MD, Deborah Swartz-Basile PhD, Carol Svatek BS, Hayder Al-Azzawi MD, Henry Pitt MD, FACS Medical College of Wisconsin, Milwaukee, WI INTRODUCTION: Cholesterol gallstone formation requires three factors: abnormal gallbladder motility, enhanced cholesterol crystallization, and supersaturated bile. We have reported that obese, leptindeficient and leptin-resistant mice have abnormal gallbladder emptying and altered cholesterol crystal formation. But the genetic factors related to gallbladder motility and crystal formation have not been elucidated. The aim of this study was to identify which gallbladder genes related to gallstone pathogenesis are regulated by leptin. METHODS: Twenty-four eight-week old leptin-deficient mice were fed a non-lithogenic diet for four weeks. 12 mice received daily IP saline injections while 12 received 5 g/g recombinant leptin. Gallbladder mRNA was isolated, pooled (4 saline, 4 leptin) and analyzed on murine microarray chips. RESULTS: The leptin-deficient mice receiving leptin had significant weight loss (34 vs 49g, p⬍0.001) and reduction in gallbladder volume (17 vs 42 l, p⬍0.001). They also had significant upregulation of the leptin receptor (p⬍0.01), JAK 1 (p⬍0.001), STAT 1 and