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Supplementary Tables 1-4, Figures 1-4 from Peloruside- and Laulimalide-Resistant Human Ovarian Carcinoma Cells Have βI-Tubulin Mutations and Altered Expression of βII- and βIII-Tubulin Isotypes

High mammographic density (MD) is a strong risk factor for breast cancer; however, data on the association between MD and breast cancer subtypes has been inconsistent and most studies have been conducted using data from Western women. Studies have shown that Asian women, who in general have denser breasts compared with Western women, were more likely to develop luminal B and HER2-enriched tumors. We previously reported that higher MD, assessed using Breast Imaging Reporting and Data System (BI-RADS) categories, was associated with HER2-enriched tumors among Chinese breast cancer patients, which may at least partially explain the higher incidence of HER2-enriched tumors observed in Asian women. Since BI-RADS MD measurement is categorical and known to be subjective, in this study, we aimed to re-evaluate the MD-subtype association using an automated quantitative density metric. The analysis included 1,549 women with invasive breast cancer who were diagnosed and treated in a tertiary hospital in Beijing, China. Subtypes were defined as Luminal A (n=606): ER+ and PR+, HER2-, and low Ki-67 ( Citation Format: Xiaohong R Yang, Yuan Tian, Jennifer Guida, Hela Koka, Ariane Chan, Changyuan Guo, Erni Li, Eric Tang, Hyuna Sung, Ning Lu, Nan Hu, Gretchen Gierach, Jing Li. Association between quantitative mammographic density and breast cancer subtypes among Chinese breast cancer patients [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P5-08-20.

PURPOSE: Mammographic density (MD) is a strong risk factor for breast cancer, yet its relationship with tumor characteristics is not well established, particularly in Asian populations. METHODS: MD was assessed from a total of 2001 Chinese breast cancer patients using Breast Imaging Reporting and Data System (BI-RADS) categories. Molecular subtypes were defined using immunohistochemical status on ER, PR, HER2, and Ki-67, as well as tumor grade. Multinomial logistic regression was used to test associations between MD and molecular subtype (luminal A = reference) adjusting for age, body mass index (BMI), menopausal status, parity, and nodal status. RESULTS: The mean age at diagnosis was 51.7 years (SD = 10.7) and the average BMI was 24.7 kg/m(2) (SD = 3.8). The distribution of BI-RADS categories was 7.4% A = almost entirely fat, 24.2% B = scattered fibroglandular dense, 49.4% C = heterogeneously dense, and 19.0% D = extremely dense. Compared to women with BI-RADS = A/B, women with BI-RADS = D were more likely to have HER2-enriched tumors (OR = 1.81, 95% CI 1.08–3.06, p = 0.03), regardless of menopausal status. The association was only observed in women with normal (< 25 kg/m(2)) BMI (OR = 2.43, 95% CI 1.24–4.76, p < 0.01), but not among overweight/obese women (OR: 0.98, 95% CI 0.38–2.52, p = 0.96). CONCLUSIONS: Among Chinese women with normal BMI, higher breast density was associated with HER2-enriched tumors. The results may partially explain the higher proportion of HER2+ tumors previously reported in Asian women.

BackgroundStudies investigating associations between mammographic density (MD) and breast cancer subtypes have generated mixed results. We previously showed that having extremely dense breasts was associated with the human epidermal growth factor receptor-2 (HER2)-enriched subtype in Chinese breast cancer patients.MethodsIn this study, we reevaluated the MD-subtype association in 1549 Chinese breast cancer patients, using VolparaDensity software to obtain quantitative MD measures. All statistical tests were 2-sided.ResultsCompared with women with luminal A tumors, women with luminal B/HER2- (odds ratio [OR] = 1.20, 95% confidence interval [CI] = 1.04 to 1.38; P = .01), luminal B/HER2+ (OR = 1.22, 95% CI = 1.03 to 1.46; P = .03), and HER2-enriched tumors (OR = 1.30, 95% CI = 1.06 to 1.59; P = .01) had higher fibroglandular dense volume. These associations were stronger in patients with smaller tumors (ConclusionAlthough biological mechanisms remain to be investigated, the associations for the HER2-enriched and luminal B subtypes with increasing MD may partially explain the higher prevalence of luminal B and HER2+ breast cancers previously reported in Asian women.

Higher mammographic density (MD) is a known risk factor for breast cancer (BC), however, studies examining determinants of density in Chinese populations are limited. Asian women tend to have higher MD compared to European women, and yet they have a lower overall BC incidence rate. The goal of this study was to investigate the influence of established BC risk factors on quantitatively measured MD among Chinese women. The study population included 7,351 Chinese women, who had mammograms at the National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China, between 2016 and 2017 for different indications, including but not limited to screening exam, breast pain, and suspected breast tumors. Only subjects with BI-RADS (Breast Imaging-Reporting and Data System) diagnostic classification of 1 (negative, n= 2,293), 2 (benign, n= 3,519), and 3 (probably benign, n= 1,539) were included in this analysis. VolparaDensity software was used to obtain quantitative MD measures including absolute dense breast volume and percent breast density. We conducted multivariable linear regression modeling on each density measurement to test for its associations with well-established breast cancer risk factors. The mean age of this study population was 50.1 years old (range= (25, 82)), the average body mass index (BMI) was 24.0 kg/m2 (range= (14.2, 48.9)), and about 46% of women were postmenopausal. The mean absolute dense and percent dense volumes were 58.4 (SD= 32.1) and 14.8 (SD= 7.1, equivalent to Volpara Density Grade= 3), respectively. Associations between quantitative MD measures and BC risk factors did not vary significantly across the three BI-RADS diagnostic classes and we therefore combined these patients in subsequent analyses. Most risk factors we examined showed similar associations with dense volume and percent density. For example, both dense volume and percent density were inversely associated with age, parity, longer duration (> 1 year) of breastfeeding, and postmenopausal status, while they were positively associated with age at menopause. In contrast, the associations with age at menarche and BMI showed opposite directions for dense volume and percent density. While increasing BMI was positively associated with dense volume, it was inversely associated with percent density. Age at menarche, however, was inversely correlated with dense volume, but positively associated with percent density. When stratifying by menopausal status, the associations for age, BMI, and parity were similar among pre- and post-menopausal women, while the associations for age at first birth and breastfeeding were stronger among postmenopausal women. In general, observed breast cancer risk factor associations with quantitative breast density metrics in this Chinese population are similar to those previously reported in Western women. We did not observe striking differences in associations of breast density with reproductive factors and BMI among Asian women in this large analysis based on quantitative density measurements. Other risk factors such as genetic susceptibility loci associated with MD should be examined in large Asian studies to further our understanding on how MD influences breast cancer risk in Asian populations. Citation Format: Hela Koka, Yuan Tian, Deng Lu, Kai Yu, Er-Ni Li, Changyuan Guo, Bin Zhu, Jennifer L Guida, Ariane Chan, Nan Hu, Ning Lu, Gretchen Gierach, Jing Li, Xiaohong R Yang. Mammographic density in relation to breast cancer risk factors among Chinese women [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P3-01-26.

Chemotherapeutic agents can induce accelerated senescence in tumor cells, an irreversible state of cell cycle arrest. Paclitaxel, a microtubule-stabilizing agent used to treat solid tumors of the breast, ovary, and lung and discodermolide, another stabilizing agent from a marine sponge, induce senescence in cultured cancer cells. The aim of this study was to determine if the microtubule-stabilizing agent peloruside A, a polyketide natural product from a marine sponge, can induce accelerated senescence in a breast cancer cell line MCF7. Doxorubicin, a DNA-damaging agent, paclitaxel, and discodermolide were used as positive controls. Senescence-associated-β-galactosidase activity was increased by peloruside A, similar to paclitaxel, discodermolde, and doxorubicin, with a potency heirarchy of doxorubicin > paclitaxel > discodermolide > peloruside, based on IC25 concentrations that inhibit proliferation. Clonogenic survival was significantly decreased by peloruside A, similar to doxorubicin and the two other microtubule-stabilizing agents. The tumor suppressor protein p53 increased after treatment, whereas pRb decreased in response to all four compounds. It was concluded that in addition to apoptosis, peloruside A causes accelerated senescence in a subpopulation of MCF7 cells that contributes to its potential anticancer activity in a breast cancer cell line.

The goal of augmented intelligence is to increase efficiency and effectiveness in practice. To achieve this, augmented intelligence technologies are being asked to perform a range of tasks, from simple to complex and quantitative. The development of these systems is increasingly important as screening becomes more personalized. This article will provide an overview of augmented intelligence in a variety of breast imaging applications.The incorporation of AI and ML techniques in breast imaging provides important new tools that will deliver ways to "sharpen" trusted familiar tools (so-called "augmented intelligence") to support radiologists, not replace them. The first wave of medical imaging systems based on AI and ML has primarily used ML to fix the values of key imaging parameters to be adapted to the individual as part of personalized medicine. Artificial intelligence is the new tool in the radiologist's arsenal but will never replace the human qualities that are important in medicine-intellectual curiosity, passion, and drive.

Mammographic breast density (MBD) has been proven to be an important risk factor for breast cancer and an important determinant of mammographic screening performance. The measurement of density has changed dramatically since its inception. Initial qualitative measurement methods have been found to have limited consistency between readers, and in regards to breast cancer risk. Following the introduction of full-field digital mammography, more sophisticated measurement methodology is now possible. Automated computer-based density measurements can provide consistent, reproducible, and objective results. In this review paper, we describe various methods currently available to assess MBD, and provide a discussion on the clinical utility of such methods for breast cancer screening.

Several analogues of the fungal natural product aigialomycin D (AmD) have been synthesised. These include the stereoisomer 5′ R ,6′ S -AmD, 2,4-di-deoxyAmD, 1′,2′,7′,8′-tetrahydroAmD and a 15-membered macrocyclic sulfone. Growth inhibitory activities of these compounds against the HL-60 leukaemic cell line were measured. The ring-expanded sulfone and tetrahydro-analogue were found to have similar IC 50 values to the natural product, whereas the 5′ R ,6′ S -stereoisomer was inactive. Energy minimisation of AmD and the synthesised analogues resulted in a range of lowest energy conformers, from planar, open arrangements of the macrocycle in AmD and tetrahydroAmD to bent, L-shaped structures for the sulfone. The synthesis of methyl orsellinate was investigated and optimised as part of this work. A stereodivergent route to both enantiomers of the diol fragment from d -ribose was also achieved.

The purposes of this study were to compare BI-RADS density categories with quantitative volumetric breast density (VBD) for the reporting of mammographic sensitivity and to identify which patient factors are most predictive of a diagnosis of interval cancer of the breast versus screen-detected cancer.This retrospective study included screen-detected cancers (n = 652) and interval cancers (n = 119) identified between January 2009 and December 2012. Multivariate logistic regression analysis was used to determine which patient factors are predictive of a diagnosis of interval cancer. Sensitivity (screen-detected cancer / [screen-detected cancer + interval cancer]) was determined with the BI-RADS 4th edition density categories and an automated equivalent density grade obtained with a proprietary tool. Sensitivity changes within automated density grade categories were investigated by use of quantitative thresholds at the midpoints of each category.In univariate analysis, age, menopausal status, and breast density were associated with a diagnosis of interval cancer. Of these risk factors, breast density was the only independent factor whether it was assessed by visual BI-RADS category (odds ratio, 3.54; 95% CI, 1.55-8.10), automated density grade (odds ratio, 4.68; 95% CI, 2.26-9.67), or VBD (odds ratio, 4.51; 95% CI, 1.92-10.61). Sensitivity decreased consistently across increasing automated density grade categories from fatty to extremely dense (95%, 89%, 83%, 65%) and less so for visual BI-RADS (82%, 90%, 84%, 66%). Further dichotomization with VBD cutoffs showed a striking linear relation between VBD and sensitivity (RIn this study, breast density was the only risk factor significantly associated with a diagnosis of interval cancer versus screen-detected cancer. Quantitative VBD captures the potential masking risk of breast density more precisely than does the widely used visual BI-RADS density classification system.

To ensure proper chromosome segregation, mitosis is tightly regulated by the spindle assembly checkpoint (SAC). Low concentrations of microtubule-stabilizing agents can induce aneuploid populations of cells in the absence of G2/M block, suggesting pertubation of the spindle checkpoint. We investigated the effects of peloruside A, a microtubule-stabilizing agent, on expression levels of several key cell cycle proteins, MAD2, BUBR1, p55CDC and cyclin B1. Synchronized 1A9 ovarian carcinoma cells were allowed to progress through the cell cycle in the presence or absence of peloruside A. Co-immunoprecipitation and Western blotting were used to probe the cell cycle kinetics of MAD2 and BUBR1 dissociation from p55CDC. Using confocal microscopy, we investigated whether premature dissociation of MAD2 and BUBR1 at low (40 nM) but not high (100 nM) concentrations of peloruside A was caused by defects in the attachment of chromosomes to the mitotic spindle. An increased frequency of polar chromosomes was observed at low concentrations of peloruside A, suggesting that an increased frequency of pseudo-metaphase cells, which are not detected by the spindle assembly checkpoint, may be underlying the induction of aneuploidy.

Peloruside A (PelA), a novel microtubule-stabilizing agent and potential anti-cancer drug, isolated from the marine sponge Mycale hentscheli, binds to a distinct, non-taxoid binding site on tubulin. Using live-cell confocal microscopy, the effects of PelA on microtubule dynamics were quantified in a human breast adenocarcinoma cell line (MCF7) stably expressing GFP-α-tubulin. Changes in microtubule length were tracked over time in cells treated with PelA concentrations ranging from 3.8-100 nM. As with other microtubule-targeting drugs like paclitaxel and epothilone B, microtubule dynamics were suppressed in a concentration-dependent manner. At the PelA IC₅₀ concentrations for cell proliferation (3.8 nM) and G₂/M block (25 nM), PelA inhibited dynamicity by 23% and 45%, respectively. At 25 nM PelA, effects included a 24% and 41% reduction in average growth rate and growth length, respectively. Additionally, the total time spent in pause increased by 53% and coincided with a 36% reduction in the average amount of time spent growing. Rescue and catastrophe frequencies were not significantly affected by PelA, except for length-based catastrophe (67% increase). The results provide further insight into PelA's unique mode of stabilization and contribute to our understanding of how microtubule-targeting agents exert their anti-mitotic effects.

Peloruside A is a microtubule-stabilizing agent that is currently under investigation as a potential anticancer agent. Peloruside A binds to a site on β-tubulin that is distinct to that of the taxanes (paclitaxel and docetaxel) and the epothilones. An attractive clinical quality of microtubule-stabilizing agents is their ability to target multiple mechanisms of tumour growth. In addition to inducing tumour cell apoptosis by arresting cells in mitosis, microtubule-stabilizing agents also inhibit angiogenesis, a process needed by tumor cells for growth and metastasis. In this study, the effects of peloruside A on endothelial cell processes important for angiogenesis were assessed in comparison to docetaxel. Both peloruside A and docetaxel potently inhibited the proliferation of human umbilical vein endothelial cells, with IC50 values of 1.4 and 1.7 nM, respectively. Peloruside also potently blocked endothelial cell migration during wound closure and the three-dimensional organization of the endothelial cells into capillary-like tubes. In the wound scratch assay, peloruside A inhibited wound recovery with an IC50 of 6.3 nM after 18 h. Docetaxel was approximately 3-fold more potent than peloruside A. The number of capillary-like tubes that formed after 16 h culture in Matrigel™ was also inhibited in a dose-dependent manner with an IC50 of 4.5 nM. Docetaxel was about 2-fold more potent than peloruside A in preventing tube formation. This inhibition of endothelial cell function occurred at relatively non-cytotoxic concentrations over the 16–18 h incubations for both stabilizing agents, suggesting that anti-angiogenic effects are likely to occur before therapeutically relevant doses begin to inhibit tumor growth or adverse side effects develop.

Item does not contain fulltext BACKGROUND: A lack of consistent guidelines regarding mammographic compression has led to wide variation in its technical execution. Breast compression is accomplished by means of a compression paddle, resulting in a certain contact area between the paddle and the breast. This procedure is associated with varying levels of discomfort or pain. On current mammography systems, the only mechanical parameter available in estimating the degree of compression is the physical entity of force (daN). Recently, researchers have suggested that pressure (kPa), resulting from a specific force divided by contact area on a breast, might be a more appropriate parameter for standardization. Software has now become available which enables device-independent cross-comparisons of key mammographic metrics, such as applied compression pressure (force divided by contact area), breast density and radiation dose, between patient populations. PURPOSE: To compare the current compression practice in mammography between different imaging sites in the Netherlands and the United States from a mechanical point of view, and to investigate whether the compression protocols in these countries can be improved by standardization of pressure (kPa) as an objective mechanical parameter. MATERIALS AND METHODS: We retrospectively studied the available parameters of a set of 37,518 mammographic compressions (9188 women) from the Dutch national breast cancer screening programme (NL data set) and of another set of 7171 compressions (1851 women) from a breast imaging centre in Pittsburgh, PA (US data set). Both sets were processed using VolparaAnalytics and VolparaDensity to obtain the applied average force, pressure, breast thickness, breast volume, breast density and average glandular dose (AGD) as a function of the size of the contact area between the breast and the paddle. RESULTS: On average, the forces and pressures applied in the NL data set were significantly higher than in the US data set. The relative standard deviation was larger in the US data set than in the NL data set. Breasts were compressed with a force in the high range of >15 daN for 31.1% and >20 kPa for 12.3% of the NL data set versus, respectively, 1.5% and 1.7% of the US data set. In the low range we encountered compressions with a pressure of

Polyglutamylation of tubulin and other non-tubulin substrates is a reversible posttranslational modification brought about by tubulin tyrosine-like ligases. Altered polyglutamylation is linked to tumorigenesis and resistance to chemotherapeutic drugs that target the microtubule, and therefore is a potential pharmacological target in cancer therapy. Despite the large amount of research focused on the development of anticancer agents, only a small number of well-characterized inhibitors of polyglutamylases have been identified, including the phosphinic acid-based inhibitors of Ttll7. In this minireview, we summarize the role of polyglutamylation in cancer, and draw attention to the largely unexplored area of polyglutamylase inhibition in the treatment of cancer.

Paclitaxel (Taxol®), a drug used to treat solid tumors of the breast, ovary and lung, stabilizes microtubules and arrests cells in G(2)/M of the cell cycle. Using two-dimensional differential in-gel electrophoresis (DIGE), we examined the proteomic response of a human HL-60 promyeloid leukemic cell line to paclitaxel. Our intention was to compare the effects of paclitaxel to those of a new-generation microtubule-stabilizing agent, peloruside A, investigated in an earlier study. In response to 100 nM paclitaxel treatment for 24 h, 21 identified proteins changed in abundance, with 13 increases and 8 decreases. In addition, 21 other unidentified proteins were also changed by treatment with paclitaxel. Using Western blotting, the transcription factor c-Myc was shown to be reduced in abundance by both drugs. Our results showed both differences and similarities at the single protein level between paclitaxel and peloruside A, although the same general classes of proteins: cytoskeletal, nucleic acid binding, stress, and apoptotic proteins, changed following exposure. The proteomic response to paclitaxel was more extensive than the response to an equipotent dose of peloruside A.

Peloruside A and laulimalide are potent microtubule-stabilizing natural products with a mechanism of action similar to that of paclitaxel. However, the binding site of peloruside A and laulimalide on tubulin remains poorly understood. Drug resistance in anticancer treatment is a serious problem. We developed peloruside A- and laulimalide-resistant cell lines by selecting 1A9 human ovarian carcinoma cells that were able to grow in the presence of one of these agents. The 1A9-laulimalide resistant cells (L4) were 39-fold resistant to the selecting agent and 39-fold cross-resistant to peloruside A, whereas the 1A9-peloruside A resistant cells (R1) were 6-fold resistant to the selecting agent while they remained sensitive to laulimalide. Neither cell line showed resistance to paclitaxel or other drugs that bind to the taxoid site on β-tubulin nor was there resistance to microtubule-destabilizing drugs. The resistant cells exhibited impaired peloruside A/laulimalide-induced tubulin polymerization and impaired mitotic arrest. Tubulin mutations were found in the βI-tubulin isotype, R306H or R306C for L4 and A296T for R1 cells. This is the first cell-based evidence to support a β-tubulin–binding site for peloruside A and laulimalide. To determine whether the different resistance phenotypes of the cells were attributable to any other tubulin alterations, the β-tubulin isotype composition of the cells was examined. Increased expression of βII- and βIII-tubulin was observed in L4 cells only. These results provide insight into how alterations in tubulin lead to unique resistance profiles for two drugs, peloruside A and laulimalide, that have a similar mode of action. Mol Cancer Ther; 10(8); 1419–29. ©2011 AACR.

Microtubule-stabilizing agents are an important class of anticancer compounds. Peloruside A and laulimalide bind to a different site on the microtubule to taxoid site drugs such as paclitaxel (Taxol(®)), docetaxel (Taxotere(®)), ixabepilone (Ixempra(®)), the epothilones, and discodermolide. The purpose of this study was to examine the synergistic interactions of these drugs when given in combination in relation to the differences in their binding sites on the microtubule.Human ovarian carcinoma cells (1A9 cells) and murine T cells were treated with different combinations of microtubule-stabilizing or destabilizing agents. The compounds were given individually and in combination, and the antiproliferative activity was assessed to calculate a combination index (CI) from the equation: CI = D(1)/Dx(1) + D(2)/Dx(2) in which D(1) and D(2) are the concentrations of drug 1 and drug 2 that when given together give the same response as drug 1 and 2 alone (Dx(1) and Dx(2)). Thus, a CI value of less than 1.0 indicates a synergistic effect between the two drugs in which the response to the two drugs given together is greater than the additive response of the two drugs if given on their own.As anticipated from previous in vitro studies, peloruside A and laulimalide did not synergize with each other. They also failed to synergize with the microtubule-destabilizing agents vinblastine and 2-methoxyestradiol. Peloruside A and laulimalide did, however, synergize with the epothilones, as had been previously shown, but not with docetaxel or discodermolide.Combining two microtubule-targeting agents with different binding sites does not guarantee a synergistic interaction in cells, and additional factors are likely to be involved. This study highlights the importance of preclinical testing of actual combinations of drugs before proceeding into clinical trials.

Peloruside B (2), a natural congener of peloruside A (1), was isolated in sub-milligram quantities from the New Zealand marine sponge Mycale hentscheli. Peloruside B promotes microtubule polymerization and arrests cells in the G(2)/M phase of mitosis similar to paclitaxel, and its bioactivity was comparable to that of peloruside A. NMR-directed isolation, structure elucidation, structure confirmation by total synthesis, and bioactivity of peloruside B are described in this article. The synthesis features Sharpless dihydroxylation, Brown's asymmetric allylboration reaction, reductive aldol coupling, Yamaguchi macrolactonization, and selective methylation.

Peloruside B (2), a natural congener of peloruside A (1), was isolated in sub-milligram quantities from the New Zealand marine sponge Mycale hentscheli. Peloruside B promotes microtubule polymerization and arrests cells in the G(2)/M phase of mitosis similar to paclitaxel, and its bioactivity was comparable to that of peloruside A. NMR-directed isolation, structure elucidation, structure confirmation by total synthesis, and bioactivity of peloruside B are described in this article. The synthesis features Sharpless dihydroxylation, Brown's asymmetric allylboration reaction, reductive aldol coupling, Yamaguchi macrolactonization, and selective methylation.

Peloruside A (PelA) is a novel secondary metabolite isolated from the New Zealand marine sponge Mycale hentscheli. It is a potent microtubule-stabilizer and binds to a distinct site on -tubulin compared to the widely used anti-cancer drug paclitaxel. PelA has clear potential benefits over paclitaxel, including increased solubility, reduced sensitivity to multiple drug resistance in cancer cell lines, and improved tolerability and efficacy in non-small cell lung cancer xenografts in mice. Previous chemical genetics studies have indicated that MAD2 (mitotic arrest deficiency-2), a spindle-assembly checkpoint (SAC) protein, could potentially be a secondary target of PelA. Premature dissociation of MAD2 from the SAC protein, p55CDC, has been implicated in the induction of aneuploidy in response to low concentrations of paclitaxel; thus, we sought to validate and investigate MAD2 as an additional target of PelA. Flow cytometric analysis of 1A9 (human ovarian carcinoma) cells confirmed that low concentrations of PelA (< 40 nM) induced chromosome mis-segregation in up to 15% of cells. Western blot analyses detected minimal changes in the abundance levels of MAD2, BUBR1 and p55CDC at both high (100 nM) and low (40 nM) PelA concentrations. However, differences in the association between MAD2 and BUBR1 with p55CDC were observed after cells were released from synchronization and assessed by co-immunoprecipitation. The mitotic arrest seen at 100 nM PelA was accompanied by a sustained formation of MAD2/p55CDC and BUBR1/p55CDC complexes; whereas, 40 nM PelA resulted in a similar dissociation profile of these complexes compared to controls. Given that PelA-induced aneuploidy coincides with G2-M progression, it is likely that dissociation of these complexes occurs despite aberrant microtubule-kinetochore attachments. Studies are currently underway to determine whether this increase in aneuploidy is due to the presence of erroneous attachments that are not detected by the SAC, such as merotelically oriented chromosomes. Aneuploidy is important because of its link to apoptosis and cancer cell death, and these experiments will help to elucidate some of the underlying signalling pathways. Additionally, these studies improve our understanding of the molecular actions of PelA during the cell cycle and highlight the importance of characterizing the concentration-dependent effects of drug interactions. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr C217.