Targeting microtubule stabilization to reduce breast tumor metastasis. Breast tumor cells metastasize to distant organs through non-adherent microenvironments, such as the bloodstream or lymphatics. However, very little is known about the dynamic behavior and drug responses of non-adherent tumor cells, due to the challenges of imaging non-adherent cells without blurring from cell drift. The PI's lab discovered unique microtentacles (McTNs) on the surface of non-adherent tumor cells that promote the aggregation and retention of circulating tumor cells (CTCs) in the lung capillaries of living mice. This revised study will test the hypothesis that therapeutic targeting of 2 microtubule stabilization mechanisms (detyrosination, acetylation) can provide a novel strategy to suppress tumor metastasis and move beyond the current non-selective MT-targeting drugs that are aimed at tumor cell growth. Predictions of this hypothesis will be tested in the following specific aims with confocal and electron microscopy, whole-animal imaging of CTC metastasis and by testing drugs on patient tumor cells and CTCs.
Specific Aim 1 : Inhibit tubulin detyrosination to reduce McTNs, stem cell characteristics and metastasis. A) Alter tubulin detyrosination by targeting TTL or TCP and gauge effects on metastatic potential/EMT. B) Test efficacy of Curcumin and Parthenolide analog on EMT, stem cell characteristics and metastasis. C) Define whether microtubule-stabilizing therapies (Paclitaxel, Ixabepilone) promote metastatic potential.
Specific Aim 2 : Determine the effects of altering tubulin acetylation on metastatic phenotypes. A) Genetically alter tubulin acetylation (ATAT1, K40R, HDAC6) and gauge effects on metastasis in mice. B) Analyze tumor cell mechanics and McTN structure in cells with altered tubulin acetylation. C) Examine whether elevation of tubulin acetylation by HDAC inhibition increases metastatic potential.
Specific Aim 3 : Target microtubule stabilization mechanisms in live CTCs from mice and human patients. A) Test 2 prioritized drugs on xenografts, PDX and live patient CTCs to reduce McTNs and CTC clusters. B) Compare CTC metastasis with MT-stabilizing and MT-disrupting agents in isolation and combination. An innovative microfluidic cell tethering system will be used to rapidly determine drug responses in live patient CTCs in less than one hour. Inclusion of numerous FDA-approved therapies will increase the potential to rapidly translate the outcomes of this project to impact the clinical treatment of metastatic breast cancer.
This project will test the therapeutic efficacy of targeting tubulin detyrosination and tubulin acetylation to reduce the metastatic potential of breast tumor cells. The PI's lab has established that these tubulin stabilization mechanisms support unique microtentacles that promote reattachment of tumor cells in distant tissues during metastasis. Innovative bioengineering approaches, animal models, PDX and live patient CTCs will be used to test these mechanisms of targeting specific subsets of microtubules.
|Pratt, Stephen J P; Hernández-Ochoa, Erick O; Lee, Rachel M et al. (2018) Real-time scratch assay reveals mechanisms of early calcium signaling in breast cancer cells in response to wounding. Oncotarget 9:25008-25024|
|Bailey, Patrick C; Lee, Rachel M; Vitolo, Michele I et al. (2018) Single-Cell Tracking of Breast Cancer Cells Enables Prediction of Sphere Formation from Early Cell Divisions. iScience 8:29-39|
|Kallergi, G; Aggouraki, D; Zacharopoulou, N et al. (2018) Evaluation of ?-tubulin, detyrosinated ?-tubulin, and vimentin in CTCs: identification of the interaction between CTCs and blood cells through cytoskeletal elements. Breast Cancer Res 20:67|
|Kessler, Michael D; Pawar, Nisha R; Martin, Stuart S et al. (2018) Improving Cancer Detection and Treatment with Liquid Biopsies and ptDNA. Trends Cancer 4:643-654|
|Ory, Eleanor C; Chen, Desu; Chakrabarti, Kristi R et al. (2017) Extracting microtentacle dynamics of tumor cells in a non-adherent environment. Oncotarget 8:111567-111580|
|Ory, Eleanor C; Bhandary, Lekhana; Boggs, Amanda E et al. (2017) Analysis of microtubule growth dynamics arising from altered actin network structure and contractility in breast tumor cells. Phys Biol 14:026005|
|Adams, Daniel L; Adams, Diane K; Alpaugh, R Katherine et al. (2016) Circulating Cancer-Associated Macrophage-Like Cells Differentiate Malignant Breast Cancer and Benign Breast Conditions. Cancer Epidemiol Biomarkers Prev 25:1037-42|
|Adams, Daniel L; Alpaugh, R Katherine; Martin, Stuart S et al. (2016) Precision Microfilters as an all in one System for Multiplex Analysis of Circulating Tumor Cells. RSC Adv 6:6405-6414|
|Adams, Daniel L; Adams, Diane K; Stefansson, Steingrimur et al. (2016) Mitosis in circulating tumor cells stratifies highly aggressive breast carcinomas. Breast Cancer Res 18:44|
|Chakrabarti, Kristi R; Andorko, James I; Whipple, Rebecca A et al. (2016) Lipid tethering of breast tumor cells enables real-time imaging of free-floating cell dynamics and drug response. Oncotarget 7:10486-97|
Showing the most recent 10 out of 24 publications