Significant Reduction In Ki67 Research Articles

Thymus antibody-secreting cells possess an interferon-responsive gene signature and are comprised of a CD154(CD40L) dependent plasmablast population.

Abstract Antibody-secreting cells (ASCs) consist of short-lived plasmablasts and post-mitotic mature plasma cells. While the major function of these cells is the production of antibodies, ASCs have been shown to regulate hematopoiesis and as well as the immune response in a context dependent manner. For years, it has been known that ASCs accumulate in the thymus of individuals with autoimmune disorders such as myasthenia gravis. In these instances, thymus ASCs persist long-term and generate pathogenic autoantibodies. However, it remains unclear how these cells are produced and regulated in the thymus under healthy conditions. Understanding these basic concepts may provide clues as to the emergence of select autoimmune diseases. As such, we have investigated the mechanisms of generation and the transcriptional landscape of thymus ASCs and compared these features to their counterparts in the bone marrow and spleen. Examination of ASC numbers demonstrated an organ-specific sex bias which manifested as increased numbers in the female thymus relative to males. Flow cytometric analysis for the proliferation marker Ki-67 demonstrated the presence of Ki-67 +plasmablasts. The administration of CD154(CD40L) blocking antibodies led to significant reduction in Ki-67 +ASCs as well as Ki-67 +B cells in the thymus. As CD40 expression was nearly exclusive to thymus B cells, the effect of CD154(CD40L) blockade was most likely mediated on upstream B cells. Finally, single cell RNA-sequencing revealed that thymus ASCs possessed an interferon-responsive gene signature which was validated at the protein level. Overall, these data provide insights as to the generation and regulation of thymus ASCs which can be leveraged for future studies. This work was supported by the National Institute on Aging of the National Institutes of Health under Award Number R03AG071955.

Loss of claudin-4 reduces DNA damage repair and increases sensitivity to PARP inhibitors (228)

Objectives: Claudin-4 is aberrantly expressed in nearly 70% of all ovarian cancer tumors, and increased claudin-4 expression conveys a worse overall survival. Poly (ADP-ribose) polymerase (PARP) inhibitors are a therapeutic option for patients with ovarian cancer. PARP inhibitors exploit deficiencies in DNA damage repair (e.g., BRCA-mutation) by inducing replication stress and DNA damage. This study aimed to define the relationship between claudin-4 expression and response to PARP inhibitors. Results: Elevated claudin-4 mRNA expression is associated with ovarian cancer, DNA damage repair pathways, and resistance to platinum-based chemotherapies and PARP inhibitors. Claudin-4 knockdown significantly sensitized ovarian cancer cells to PARP inhibitors. Specifically, loss of claudin-4 reduced the 50% inhibitory concentration (IC50) of olaparib by 70% (IC50 - 378 nM to 88 nM) and rucaparib by 85% (IC50 - 189 nM to 27.5 nM). Furthermore, loss of claudin-4 expression sensitized ovarian cancer cells to several FDA-approved chemotherapies that induce DNA damage. Claudin-4 downregulation resulted in the decrease of several DNA damage repair effectors, including 53BP1 and XRCC1. Claudin-4/53BP1/XRCC1 expression was mainly confined to the tumor cells. Claudin-4 knockdown did not change homology-directed repair but reduced both non-homologous end-joining activity by 41% (p<0.001) and 53BP1 foci formation by an average of 50% (p<0.05). In the TCGA, lower claudin-4 expression correlated to increased mutational burden (Claudin-4 “High”, 45.2 mutations vs Claudin-4 “Low” 134.6 mutations; p<0.01). In 15 primary ovarian cancer tumors (12 high-grade serous, one low-grade serous, one mucinous, and one granulosa cell), “low”/null claudin-4 expressing tumors had a significant reduction in Ki67 after PARP inhibitor treatment alone (p<0.01). In contrast, “High” claudin-4 tumors only had significantly reduced Ki67 in the claudin-4 inhibitor alone (p<0.0001) and in the CMP/olaparib combination (p<0.0001). Conclusions: Claudin-4 is related to non-homologous end-joining DNA damage repair and 53BP1 foci formation. Claudin-4 inhibition in “high” claudin-4 tumors sensitized tumors to PARP inhibition. Claudin-4 expression in high-grade ovarian cancer tumors could potentially serve as both a marker of PARP inhibitor response and a target to improve PARP inhibitor response. Objectives: Claudin-4 is aberrantly expressed in nearly 70% of all ovarian cancer tumors, and increased claudin-4 expression conveys a worse overall survival. Poly (ADP-ribose) polymerase (PARP) inhibitors are a therapeutic option for patients with ovarian cancer. PARP inhibitors exploit deficiencies in DNA damage repair (e.g., BRCA-mutation) by inducing replication stress and DNA damage. This study aimed to define the relationship between claudin-4 expression and response to PARP inhibitors. Results: Elevated claudin-4 mRNA expression is associated with ovarian cancer, DNA damage repair pathways, and resistance to platinum-based chemotherapies and PARP inhibitors. Claudin-4 knockdown significantly sensitized ovarian cancer cells to PARP inhibitors. Specifically, loss of claudin-4 reduced the 50% inhibitory concentration (IC50) of olaparib by 70% (IC50 - 378 nM to 88 nM) and rucaparib by 85% (IC50 - 189 nM to 27.5 nM). Furthermore, loss of claudin-4 expression sensitized ovarian cancer cells to several FDA-approved chemotherapies that induce DNA damage. Claudin-4 downregulation resulted in the decrease of several DNA damage repair effectors, including 53BP1 and XRCC1. Claudin-4/53BP1/XRCC1 expression was mainly confined to the tumor cells. Claudin-4 knockdown did not change homology-directed repair but reduced both non-homologous end-joining activity by 41% (p<0.001) and 53BP1 foci formation by an average of 50% (p<0.05). In the TCGA, lower claudin-4 expression correlated to increased mutational burden (Claudin-4 “High”, 45.2 mutations vs Claudin-4 “Low” 134.6 mutations; p<0.01). In 15 primary ovarian cancer tumors (12 high-grade serous, one low-grade serous, one mucinous, and one granulosa cell), “low”/null claudin-4 expressing tumors had a significant reduction in Ki67 after PARP inhibitor treatment alone (p<0.01). In contrast, “High” claudin-4 tumors only had significantly reduced Ki67 in the claudin-4 inhibitor alone (p<0.0001) and in the CMP/olaparib combination (p<0.0001). Conclusions: Claudin-4 is related to non-homologous end-joining DNA damage repair and 53BP1 foci formation. Claudin-4 inhibition in “high” claudin-4 tumors sensitized tumors to PARP inhibition. Claudin-4 expression in high-grade ovarian cancer tumors could potentially serve as both a marker of PARP inhibitor response and a target to improve PARP inhibitor response.

Abstract P1-09-04: Flaxseed &amp; breast cancer: A systematic review

Abstract Background: Flaxseed has been extensively studied in in vitro and in vivo models for possible association with reducing the risk of breast cancer. Clinical trials have been limited. There has been confusion regarding whether it is safe and beneficial for breast cancer survivors to take flaxseed. Therefore, we conducted a systematic review to evaluate the existing randomized controlled trial to understand the safety and efficacy of using flaxseed for symptom control and reducing the risk of recurrence among breast cancer survivors. Patients and Methods: We conducted a literature search of English language publications in MEDLINE, EMBASE, and Cochrane from the databases’ inception to May 2021 using the keywords flaxseed, breast cancer, randomized clinical trial, and human studies. Results: We identified three randomized controlled trials (RCT) that include patients with a history of breast cancer. Sample size ranged from 24 to 178 patients and treatment duration ranged from 13 to 42 days. The primary end points were hot flash severity (one study) and tumor biomarkers (two studies). There was no statistically significant reduction in mean hot flash score in the flaxseed group compared to the placebo group. One study showed a statistically significant reduction in Ki-67 (34.2%; P=0.001) and in c-erbB2 expression (71.0%; P=0.003), and an increase in apoptosis (30.7%; P=0.007) in the group that received flaxseed, but not in the placebo group. On the contrary, another study showed no difference in Ki-67, caspase, the Ki-67/caspase ratio, and Estrogen-receptor β. Two studies reported mild adverse events as shown in Table 1. None of the studies assessed the change in estrogen level. Risk of bias assessment showed that the overall risk of bias associated with the included studies is moderate to high. The high risk of bias originates from lack of blinding of participants and personnel. Conclusion: Current clinical research in breast cancer survivors is too limited to recommend flaxseed for cancer prevention. Flaxseed appears to be safe with mild side effects, however, the effect of flaxseed on estrogen concentration has not been established. Table 1.Characteristics of included studies and risk of bias assessmentIDBreast cancer participantsSample sizeExperimentalControlOutcomesAdverse eventsRisk of biasPruthi 2012(U.S.)With breast cancer: (Yes/No)Flaxseed: 44/88Placebo: 47/90Flaxseed: 88Placebo: 90Flaxseed bar: 7.5g of flaxseed, 200 calories, one bar daily, 6 weeksPlacebo bar: 2g protein, 20% fiber, 200 calories, one bar daily, 6 weeksFrequency and severity of hot flash; hot flash interference; the Profile of Mood States; the Menopause Quality of Life Scale; the Global Impression of BenefitAbdominal distension and pain, diarrhea, flatulence, nausea, pruritus, maculopapular rash, vomitingLowThompson 2005(Canada)Flaxseed: (ER+ &amp; PR+) 17/19; (ER- OR PR-) 2/19Placebo: (ER+ &amp; PR+) 11/13; (ER- OR PR-) 2/13Flaxseed: 19Placebo: 13Flaxseed muffin: 25g flaxseed + 20.7g white wheat flour, one daily, 32 daysPlacebo muffin: 20.7g whole wheat flour+10g canola oil, one daily, 39 daysBreast cancer biomarkers including Ki-67, apoptosis, C-erbB2, ER and PR scoresAbdominal fullness and increased bowel movementsSome concernMcCann 2014(Canada)Clinical stage: Flaxseed: I 4/6; IIA 1/6; IIB 1/6;Anastrozole: I 6/7; IIA 1/7;Flaxseed/Anastrozole: I 5/6; IIA 1/6;Placebo: I 3/5; IIA 2/5Flaxseed: 6;Anastrozole: 7;Flaxseed/Anastrozole: 6;Placebo: I5;Flaxseed: 25 g/d ground flaxseed + 1/d placebo pill, 13-16 daysAnastrozole: 1 mg/d;Flaxseed/Anastrazole: 25 g/d ground flaxseed + 1 mg/d Anastrozole;Placebo: 1/d; 13-16 daysBreast cancer biomarkers including Ki-67, caspase, the Ki-67/caspase ratio, and Estrogen-receptor βNot reportedHigh Citation Format: Rui Wang, Mingxiao Yang, Iris Zhi, Ting Bao. Flaxseed &amp; breast cancer: A systematic review [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 P1-09-04.

Changes in ki-67 and cleaved caspase-3 (CC3) with short term androgen deprivation therapy (ADT) prior to prostatectomy as part of a trial in intermediate and high-risk prostate cancer (PC).

e17502 Background: Ki-67 and CC3 are markers of cellular proliferation and apoptosis and have been associated with prognosis in localized PC. Testosterone suppression promotes a time-dependent modulation of these indices. We explored changes in Ki-67 and CC3 at different timepoints in a pilot trial using short-term ADT prior to radical prostatectomy (RP). Methods: Pts with intermediate and high-risk localized PC were randomized to receive a single dose of neoadjuvant degarelix (240 mg SC) either 4 ± 1 days (Cohort A) or 7 ± 1 days (Cohort B) prior to RP. Anatomically matched tumor foci from the pre-treatment diagnostic biopsy and the prostatectomy specimens were assessed by immunohistochemistry (IHC) for Ki-67, CC3, and PTEN expression. Results: 32 pts were randomized: 15 to Cohort A and 17 to Cohort B. Ki-67 and CC3 expression in the pre- and post-treatment samples for both cohorts are summarized in the table. A significant reduction in Ki-67 and increase in CC3 expression occurred in both cohorts (all p-values &lt; 0.05). No difference was found when the absolute difference of Ki-67 expression was compared between Cohort A and Cohort B (p = 0.7). A significant reduction of Ki-67 and increase in CC3 expression occurred in pts independent of PTEN status. Conclusions: A reduction of Ki-67 and an increase in CC3 expression was achieved in both cohorts following a single dose of degarelix within 1 week of RP. Although loss of PTEN has been associated with androgen-independent progression, reduction of proliferation and increase in apoptosis levels were achieved independent of PTEN status in this small cohort. Further analyses are ongoing to correlate the pathologic findings with clinical outcomes. Clinical trial information: NCT01542021 . [Table: see text]

Abstract P4-15-05: Biomarkers of response to neoadjuvant endocrine therapy with anastrozole (Ana) alone or in combination with fulvestrant (Ful) in ER-positive (ER+) HER2-negative (HER2-) breast cancer (PACT01 trial)

Abstract Background: In recent years, several clinical trials showed that fulvestrant (Ful), alone or in combination with an aromatase inhibitor (AI), is more effective than an AI alone. PACT01 is a randomized neoadjuvant trial of Anastrazole (Ana) alone or in combination with Ful in ER+/HER2- breast cancer. Methods: Patients with newly diagnosed ER+/HER2- breast cancers, 2 cm or larger in size, were randomized to 16 weeks of Ana (1 mg orally every day) alone or in combination with Ful (500mg IM days 1, 15, 29, and every 28 days thereafter) for 16 weeks. Patients then proceeded to surgery. Tumor tissue was collected at baseline, day 28 (D28), and at the time of surgery. Primary endpoint was the reduction of Ki67 in tumor tissue between baseline and D28. Baseline and D28 samples were stained for ER, PR, HER2, and Ki67. ER and PR were scored for intensity and percentage (H-score), HER2 was scored for intensity of membrane staining; and Ki67 was scored as percentage. Data were summarized descriptively. Changes in biomarkers from baseline to D28 were calculated and compared by Wilcoxon signed rank test. Results: PACT01 trial enrolled 72 patients. Three of them did not start treatment. Baseline samples were collected from the remaining 69 patients, and D28 samples from 60 patients (5 refused, 2 withdrew, 1 lost to follow up, 1 unknown). Samples from 18 patients had no tumor (5 at baseline, 9 at D28, 4 at both). Of the 42 patients with paired samples, 20 received Ana and 22 received Ana+Ful. All cases except one were centrally confirmed to be ER+, and all were HER2-. Table 1 summarizes median expression of Ki67, ER, and PR. Both treatment regimens led to a significant reduction in Ki67 between baseline and D28. However, Ana+Ful did not reduce Ki67 more effectively than Ana alone. Ki67 was reduced to &amp;lt;10% in 60% of the Ana arm and 68% of the Ana+Ful, which was not statistically significant.PR was similarly reduced in both treatment arms. ER was significantly reduced at D28 in the Ana+Ful arm (p=0.0004) but not in the Ana alone arm. Safety profile of both treatment arms was consistent with package insert and published studies. Median expression of Ki67, ER and PR in Anastrazole and Anastrazole + Fulvestrant Arms at Baseline and Day 28ARMTimepointNKi67 (%)ER H-scorePR H-scoreAnaBaseline2024.8182.5100.3 Day 28205.6*170.025.0Ana + FluBaseline2225.6198.120.5 Day 28225.1*117.50.0* p=0.0004. Other comparisons were not stastistically significant Conclusions:In this small neoadjuvant trial, the addition of Ful to Ana did not increase Ki67 suppression at D28. This may be due to untreated primary tumors being exquisitely sensitive to Ana and that fulvestrant may not add to it. It is also possible that the effect of Ful may be noted later in the course of treatment. Further biomarker data on tissue collected at the end of treatment will be presented at the meeting. Citation Format: Dhamne S, Nagi C, Wang T, Pavlick AC, Reusser B, Schiff R, Julie N, Niravath P, Silberfein EJ, Sedgwick EL, Sepulveda KA, Gutierrez C, Hilsenbeck SG, Chang JC, Osborne CK, Rimawi MF. Biomarkers of response to neoadjuvant endocrine therapy with anastrozole (Ana) alone or in combination with fulvestrant (Ful) in ER-positive (ER+) HER2-negative (HER2-) breast cancer (PACT01 trial) [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P4-15-05.

Abstract P5-16-13: Predictive and prognostic value of Ki-67 expression in triple-negative breast cancer before and after neoadjuvant chemotherapy with weekly paclitaxel plus carboplatin

Abstract Background: Neoadjuvant chemotherapy (NCT) is one of the main treatment strategies for patients with locally advanced breast cancer. This study was designed to demonstrate the utility of a platinum-containing treatment regimen in the neoadjuvant setting and to identify the predictive or prognostic value of Ki-67 among patients with triple-negative breast cancer (TNBC) Methods: Data from 280 patients with stage II–III TNBC were collected. All patients underwent four cycles of NCT with paclitaxel (80 mg/m2) and carboplatin (AUC 2 mg*min/ml) on days 1, 8, and 15 of a 28-day cycle. Ki-67 values were evaluated in both biopsy and excision specimens. The clinical responses to NCT were evaluated based on MRI and ultrasound examinations in accordance with the response evaluation criteria in solid tumors (RECIST) version 1.1. The pathological evaluation of surgical specimens was conducted at by the Miller-Payne (MP) grading system. Logistic regression analysis was performed to identify independent predictors of pathological complete response (pCR). Survival analysis was performed to evaluate the prognostic value of various clinical and pathological variables. Results: The overall pCR rate was 33.9%. Both the categorical and linear Ki-67 were independently correlated with pCR (P&amp;lt;0.001). There were also statistically significant differences among Ki-67 categories with respect to clinical response (P&amp;lt;0.001), Miller-Payne (MP) grades (P&amp;lt;0.001), and node status (P&amp;lt;0.001). A significant reduction of Ki-67 after NCT was most likely observed in patients with a relatively better response. Among 185 patients with a residual tumor after NCT, the tumor cell proliferation significantly decreased from a median Ki-67 value of 20.0% before chemotherapy to 15% after chemotherapy. The mean absolute reductions in Ki-67 were 29.4%, 8.5%, and -10.2% in patients with an ideal response (MP 5/4), a partial response (MP 3) and a poor response (MP2/1), respectively. In the multivariate model for non-pCR patients, Ki-67 reduction presented an independent prognostic value for relapse of disease (HR=0.986, 95% CI: 0.978-0.994; P=0.001). Residual node involvement was also an independent predictor of patient outcome (HR=0.895, 95% CI: 0.355-2.259 for 1-3 nodes and HR=2.424, 95% CI: 1.048-5.608 for more than 4 nodes, using 0 nodes as a reference; P=0.002). Better survival was more frequently observed in patients with a greater reduction in the Ki-67 value and fewer involved nodes. Conclusions: The primary Ki-67 might help in further classifying TNBCs into subtypes with different responses to chemotherapy. The significant reduction of Ki-67 after treatment could indicate a favorable prognosis in non-pCR patients. Citation Format: Wang R-X, Chen S, Jin X, Shao Z-M. Predictive and prognostic value of Ki-67 expression in triple-negative breast cancer before and after neoadjuvant chemotherapy with weekly paclitaxel plus carboplatin [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P5-16-13.

Metformin as a new anti-cancer drug in adrenocortical carcinoma.

Adrenocortical carcinoma (ACC) is a rare heterogeneous malignancy with poor prognosis. Since radical surgery is the only available treatment, more specific and effective drugs are urgently required. The anti-diabetic drug metformin has been associated with a decreased cancer prevalence and mortality in several solid tumors, prompting its possible use for ACC treatment.This paper evaluates the in vitro and in vivo anti-cancer effects of metformin using the ACC cell model H295R.Metformin treatment significantly reduces cell viability and proliferation in a dose- and time-dependent manner and associates with a significant inhibition of ERK1/2 and mTOR phosphorylation/activation, as well as with stimulation of AMPK activity. Metformin also triggers the apoptotic pathway, shown by the decreased expression of Bcl-2 and HSP27, HSP60 and HSP70, and enhanced membrane exposure of annexin V, resulting in activation of caspase-3 apoptotic effector. Metformin interferes with the proliferative autocrine loop of IGF2/IGF-1R, which supports adrenal cancer growth. Finally, in the ACC xenograft mouse model, obtained by subcutaneous injection of H295R cells, metformin intraperitoneal administration inhibits tumor growth, confirmed by the significant reduction of Ki67%.Our data suggest that metformin inhibits H295R cell growth both in vitro and in vivo. Further preclinical studies are necessary to validate the potential anti-cancer effect of metformin in patients affected by ACC.

Value of Ki-67 expression in triple-negative breast cancer before and after neoadjuvant chemotherapy with weekly paclitaxel plus carboplatin.

Neoadjuvant chemotherapy (NCT) is one of the main treatment strategies for patients with locally advanced breast cancer. In this study, we focused on the predictive and prognostic value of Ki-67 in triple-negative breast cancer (TNBC) patients who received NCT. Data from 280 patients with stage II–III TNBC were collected. All patients were treated according to the same protocol with weekly paclitaxel and carboplatin. The overall pCR rate was 33.9%. Both the categorical and linear Ki-67 were independently correlated with pCR (P < 0.001). There were also statistically significant differences among Ki-67 categories with respect to clinical response (P < 0.001), Miller-Payne (MP) grades (P < 0.001), and node status (P < 0.001). A significant reduction of Ki-67 after NCT was most likely observed in patients with a relatively better response. In the multivariate model for non-pCR patients, Ki-67 reduction presented an independent prognostic value for relapse of disease (HR = 0.986, 95% CI: 0.978–0.994; P = 0.001). This study had indicated that the primary Ki-67 might help in further classifying TNBCs into subtypes with different responses to chemotherapy and a significant reduction of Ki-67 after treatment could indicate a favorable prognosis in non-pCR patients.

Anti-Tumoral Effects of Anti-Progestins in a Patient-Derived Breast Cancer Xenograft Model.

Breast cancer is a hormone-dependent disease in which estrogen signaling targeting drugs fail in about 10 % due to resistance. Strong evidences highlighted the mitogen role of progesterone, its ligands, and the corresponding progesterone receptor (PR) isoforms in mammary carcinoma. Several PR antagonists have been synthesized; however, some of them are non-selective and led to side or toxic effects. Herein, we evaluated the anti-tumor activity of a commercially available PR modulator, ulipristal acetate (UPA), and a new selective and passive PR antagonist "APR19" in a novel preclinical approach based on patient-derived breast tumor (HBCx-34) xenografted in nude mice. As opposed to P4 that slightly reduces tumor volume, UPA and APR19 treatment for 42 days led to a significant 30 % reduction in tumor weight, accompanied by a significant 40 % retardation in tumor growth upon UPA exposure while a 1.5-fold increase in necrotic areas was observed in APR19-treated tumors. Interestingly, PR expression was upregulated by a 2.5-fold factor in UPA-treated tumors while APR19 significantly reduced expression of both PR and estrogen receptor α, indicating a potential distinct molecular mechanism among PR antagonists. Cell proliferation was clearly reduced in UPA group compared to vehicle conditions, as revealed by the significant reduction in Ki-67, Cyclin D1, and proliferating cell nuclear antigen (PCNA) expression. Likewise, an increase in activated, cleaved poly(ADP-ribose) polymerase (PARP) expression was also demonstrated upon UPA exposure. Collectively, our findings provide direct in vivo evidence for anti-progestin-mediated control of human breast cancer growth, given their anti-proliferative and pro-apoptotic activities, supporting a potential role in breast cancer therapy.

Cell proliferation is reduced in the hippocampus in schizophrenia.

Objective:The molecular and cellular basis of structural and functional abnormalities of the hippocampus found in schizophrenia is currently unclear. Postnatal neurogenesis contributes to hippocampal function in animal models and is correlated with hippocampal volume in primates. Reduced hippocampal cell proliferation has been previously reported in schizophrenia, which may contribute to hippocampal dysfunction.Method:We measured the cell proliferation marker, Ki67, in post-mortem hippocampal tissue from patients with schizophrenia (n = 10) and matched controls (n = 16). Ki67-labelled cells were counted within the dentate gyrus and hilus on sections taken from the anterior hippocampus.Results:We replicated the finding of a significant reduction in Ki67+ cells/mm2 in schizophrenia cases compared to controls (t24 = 2.1, p = 0.023). In our relatively small sample, we did not find a relationship between Ki67+ cells and age overall, or between Ki67 + cells and duration of illness or antipsychotic treatment in people with schizophrenia.Conclusion:Our results confirm that reduced hippocampal cell proliferation may be present in schizophrenia. Restoring hippocampal neurogenesis may be a potential therapeutic target for the treatment of hippocampal dysfunction in schizophrenia.

Pure anti-tumor effect of zoledronic acid in naïve bone-only metastatic and locally advanced breast cancer: proof from the “biological window therapy”

The study investigated the anti-tumour effect of zoledronic acid (ZA) administered alone in a biological window therapy in naïve bone-only metastatic and locally advanced breast cancer (LABC) patients. 33 patients with LABC (Group 1) and 20 patients with a first diagnosis of bone metastasis only (Group 2) received 4 mg single dose of ZA, 14 days (biological window) before starting any treatment. In Group 1, Ki67, CD34, p53/bcl-2 and caspase 3 expression along with the adenosine triphosphate (ATP) levels and RNA disruption index were evaluated as markers of tumor growth in tumour specimens obtained before and after ZA administration (basal, day 14). In Group 2, the total enumeration of circulating tumour cells (CTCs), and of M30+ve CTCs along with the soluble marker of cell death (M30/M65) were carried-out as markers of tumor dissemination at baseline, at 48 h and day 14th. In Group 1, there was a significant reduction in Ki67, CD34, bcl-2 expression after 14 days ZA based-treatment (p = 0.0032; p = 0.0001, p < 0.00001 respectively). ZA showed a significant increase of RNA disruption (p < 0.0076). In Group 2, we observed a significant reduction of CTCs number after 48 h (p = 0.0012), followed by a significant rebound at 14 days (p = 0.012). The apoptotic CTCs/M30+ve and M65 levels significantly increased under treatment (p = 0.018 and p = 0.039 respectively) after drug administration when compared to the baseline. These results are the first prospective in vivo data showing the direct pure anti-tumour effect (either on the tumour cell or on CTCs) of ZA.

An open-label study of lapatinib in women with HER-2-negative early breast cancer: the lapatinib pre-surgical study (LPS study)

BackgroundThis phase II, open-label, multicentre study aimed to evaluate changes in cell proliferation and biomarkers, as well as efficacy of lapatinib in treatment-naïve patients with HER-2-negative primary breast cancer. Patients and methodsPatients received 1500 mg lapatinib for 28–42 days before surgery with repeat biopsies and measurements. The primary end point was inhibition of cell proliferation measured by Ki67; the secondary end points included clinical response, adverse events and changes in FOXO3a, FOXM1, p-AKT and HER-3. ResultsOverall, there was no significant reduction in Ki67 with treatment (assessment carried out in 28 of 31 subjects enrolled). However, four patients (14%) showed a reduction in Ki67 ≥50%. Four of 25 patients (16%) had a partial response to treatment judged by sequential ultrasound measurements. Response, in terms of either Ki67 or ultrasound, did not relate to changes in any biomarker assessed at baseline, including the estrogen receptor (ER) and epidermal growth factor receptor (EGFR). However, all four clinical responders were HER-3 positive, as were three of four Ki67 responders. ConclusionsOverall, a pre-surgical course of lapatinib monotherapy had little effect on this group of patients; however, in subsets of patients, especially those with HER-3-positive tumors, we observed either reduction in proliferation (Ki67) or tumor size; EGFR/ER status had no impact.

Cytotoxic activity of the histone deacetylase inhibitor panobinostat (LBH589) in anaplastic thyroid cancer in vitro and in vivo

Anaplastic thyroid carcinoma (ATC) has a rapidly fatal clinical course, being resistant to multimodal treatments. Microtubules, α/β tubulin heterodimers, are crucial in cell signaling, division and mitosis and are among the most successful targets for anticancer therapy. Panobinostat (LBH589) is a potent deacetylase inhibitor acting both on histones and nonhistonic proteins, including α-tubulin. In vitro LBH589, evaluated in three ATC cell lines (BHT-101, CAL-62 and 8305C), resulted in impairment of cell viability, inhibition of colony formation, cell cycle arrest and apoptosis induction. Mechanistically, we showed that LBH589 not only affected the expression of p21 and cyclin D1, but markedly determined microtubule stabilization as evidenced by tubulin acetylation and increased tubulin polymerization. In a SCID xenograft model implanted with CAL-62 cells, the cytotoxic properties of LBH589 were confirmed. The drug at the dose of 20 mg/kg significantly impaired tumor growth (final tumor volume 2.5-fold smaller than in untreated animals); at this dose, no relevant side effects were observed. In tumors of treated animals, a significant reduction of Ki67, which was negatively correlated with tubulin acetylation, was observed. Moreover, acetyl-tubulin levels negatively correlated with tumor volume at sacrifice, reinforcing the opinion that tubulin acetylation has a role in the inhibition of tumor growth. In conclusion, LBH589, acting on both histones and nonhistonic proteins in anaplastic thyroid cancer, appears to be a promising therapeutic agent for the treatment of this kind of cancer which is known not to respond to conventional therapy.

Abstract 3670: A randomized, placebo-controlled, phase II, presurgical biomarker trial of celecoxib or exemestane in postmenopausal breast cancer patients

Abstract Background Treatment of breast cancer (BC) before surgery provides a window of opportunity to screen the activity and mechanism of action of selected agents. Exemestane (Exe) is an irreversible inactivator of aromatase, the enzyme converting androgens in estrogens. Celecoxib (Cel) is a selective inhibitor of cyclooxygenase-2 (Cox-2), the enzyme involved in the production of prostaglandins that play a key role in tumor proliferation. We conducted a double-blind, placebo-controlled, randomized trial to evaluate the activity and efficacy of Cel and Exe given for 6 weeks before surgery. The main endpoint biomarker was Ki-67 labeling index (LI), a well known marker of tumor proliferation. Secondary endpoints were the modulation of biomarkers associated to BC and cardiovascular disease. Methods Postmenopausal women with histologically-confirmed ER positive BC; stage T1-2, N0-1, M0 were randomly assigned to either Exe 25 mg/d (n=50), or Cel 800 mg/d (n=50) or placebo (n=25) for 6 weeks before surgery. Fasting blood samples were withdrawn at baseline and the day before surgery. Baseline median age was 62, 65, 63 years, and median BMI was 26.56, 24.89, 26.67 in the Exe, Cel and placebo group, respectively. Changes in biomarkers were analyzed through an ANCOVA model adjusting for baseline values. Results Compared to baseline, Exe showed a 10% statistically significant reduction in KI-67% LI (95%CI; -12.52, -7.75) and a 26% reduction in progesterone receptor (PgR) expression (95% CI; -32.67, -18.62) while in the Cel group, we observed a 5% decrease in PgR expression (95%CI: -12.08; 1.84). No change was observed in ER expression. In the placebo arm Ki-67% LI, ER and PgR expression did not change significantly. Compared to baseline, Exe significantly increased testosterone levels by 0.25 ng/mL (95%CI; 0.21, 0.28), while no modulation was observed in the other two arms: Least square means of changes in testosterone were -0.005 (95%CI:-0.05, 0.04) in the placebo arm and -0.001 (95%CI:-0.03, 0.03) in the Cel arm (P for differences between treatment arms&amp;lt;0.0001), respectively. As regards high-sensitivity C-reactive protein levels, no change was observed in any arm. Exe induced a statistically significant -10.87 (95%CI:-17.32, -4.41) decrease in total cholesterol and -6.61(95%CI: -9.63,-3.59) in HDL cholesterol. Triglycerides levels were unchanged. Conversely, Cel induced a borderline statistically significant reduction of antithrombin III levels (P among active treatments =.08) while no difference was observed for fibrinogen levels. Conclusions Our study confirms the anti-proliferative effects of Exe on BC, together with a favourable effect on lipid profile. Further investigations, including the modulation of markers of bone metabolism and the activity of Cox-2 will be conducted to better elucidate the mechanism of action of both drugs in BC prevention. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3670. doi:10.1158/1538-7445.AM2011-3670

Designing adjuvant treatment based on biological measurements in the neoadjuvant setting.

For many years, one of the main aims of neoadjuvant therapy has been to identify short term endpoints that will predict for long-term outcome in adjuvant treatment. Th e advantages are obvious: in contrast to adjuvant trials, neoadjuvant trials require hundreds rather than thousands of patients, are very much less expensive to run and produce outcome data many years earlier. A key primary endpoint for neoadjuvant trials has been objective clinical response and this still remains the case [1]. Nevertheless, major studies have frequently shown that clinical response, including complete clinical remission, does not predict for long-term outcome. For example, in the National Surgical Adjuvant Breast and Bowel Project (NSABP) B-18 trial no diff erence in outcome was seen between patients who achieved clinical response versus those who did not [2]. Likewise, in the NSABP B-27 trial in which 2,411 patients were randomised to neoadjuvant adriamycin/cyclophospha mide (AC) versus the same treatment followed by sequential docetaxel, the complete response rate was 64% for the sequential, docetaxel-containing arm compared with 40% for the AC arm alone (P < 0.001), but no signifi cant diff erence was seen in long-term survival [3,4]. Likewise, for neoadjuvant endocrine therapy, the IMPACT (Immediate Preopera tive Anastrozole, Tamoxifen, or Combined With Tamoxifen) trial compared neo adjuvant anastrazole versus tamoxifen versus the combination and was the neoadjuvant equivalent of the ATAC (Arimidex, Tamoxifen, Alone or in Combination) adjuvant trial. No signifi cant diff erence was seen in response rate between the three arms of IMPACT [5] and this failed, therefore, to correlate with the long-term disease-free survival advantage of anastrazole in ATAC. Likewise, in the IMPACT trial a large diff erence was seen in response rates for patients with human epidermal growth factor receptor 2 (HER2)-positive tumours in favour of anastrazole (7 of 12 responders; 58%) compared with tamoxifen (2 of 9 responders; 22%) and yet no selective disease-free survival advantage for anastrazole over tamoxifen in patients with HER2-positive tumours was subsequently seen in ATAC [6]. A possible explana tion for this is that it is entirely possible that tumour growth rate may be slowed by neoadjuvant therapy (as measured, for example, by a reduction in proliferation factor Ki67) without a formal clinical response being achieved but with a consequent gain in relapse free survival. In the IMPACT trial, for example, only 37% of tumours achieved an objective clinical response rate to anastrazole whereas 75% had a signifi cant reduction in Ki67 [7].

Abstract 951: Inhibition of lung tumorigenesis by oral consumption of pomegranate fruit extract

Abstract Lung cancer is the most common cause of cancer mortality worldwide and continues to be a major health problem, exceeding the combined mortality rates of colorectal, breast and prostate cancers. It has proven difficult to control lung cancer with conventional therapeutic and surgical approaches, and generally the prognosis is poor with an overall five-year survival rate of 10-14% in the USA. A competent approach to reduce the mortality from lung cancer is the early detection of premalignant lesions as the survival of early stage lung cancer patients is much better as compared to patients with advanced cancers. The current mainstays of lung cancer therapy are surgical resection, radiation ablation and systemic chemotherapy with considerable toxicity. However, it appears unlikely that these therapies can cause striking improvements in near future. Thus, effective approaches to the prevention of lung cancer need to be developed, especially efforts targeted to the progression of preneoplastic and benign neoplastic lesions to more malignant lung tumors. The pomegranate (Punica granatum; Punicaceae) fruit has been used for centuries in ancient cultures for its medicinal purposes and we have been defining the chemopreventive effects of pomegranate fruit extract (PFE). Based on MALDI-TOF MS analysis, PFE was found to contain anthocyanins (such as delphinidin, cyanidin and pelargonidin) and hydrolyzable tannins (such as punicalin, pedunculagin, punicalagin, gallagic and ellagic acid esters of glucose). We have earlier shown that PFE inhibits prosurvival signaling pathways in human lung carcinoma A549 cells and tumor growth in athymic nude mice (Carcinogenesis 2007;28:163-73) and A/J mice (Cancer Res 2007;67:3475-82). In this study, the chemopreventive activity of PFE during progression of lung adenomas was investigated in two mouse tumor protocols. Female A/J mice were treated with 0.2% PFE in drinking water (w/v, equivalent of human consumption of juice derived from two fruits) till the termination of the experiment. One week after treatment with PFE, mice were given a single i.p. injection of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) at dose of 100 mg/kg body weight or urethane at dose of 1 mg/g body weight. Mice were sacrificed at 16 weeks after NNK treatment and 20 weeks after treated with urethane. PFE administration significantly reduced the lung tumor multiplicity by 47% in mice treated with NNK and 52% in urethane-treated mice. Markers of cell proliferation, apoptosis and angiogenesis were studied by immunohistochemical staining. Treatment with PFE caused significant reduction in Ki-67, proliferating cell nuclear antigen (PCNA), caspase-3, CD-31 and vascular endothelial growth factor (VEGF) in lungs of mice treated with NNK and urethane. Thus, our results show that PFE can act as a chemopreventive agent in smokers and ex-smokers with early lung lesions. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 951.

The Novel Aurora Kinase Inhibitor ENMD-2076 Has Potent Single Agent Activity against Multiple Myeloma (MM) in Vitro and in Vivo, and Shows Synergistic Activity in Combination with Lenalidomide

ENMD-2076 is a novel, orally-active molecule that has been shown to have significant activity against Aurora A kinase as well as multiple receptor tyrosine kinases (RTK). We investigated the single agent activity of ENMD-2076 against MM cells in vitro and in vivo, and in combination with lenalidomide. ENMD-2076 free base showed significant cytotoxicity against MM cells with a mean LC50 of 3.84±0.86 μM at 48 hours in vitro. Cytotoxicity was associated with cleavage of caspase 3, 8, 9 and PARP, and loss of mitochondrial membrane potential as early as 6 hours. ENMD-2076 free base inhibited c-kit, FGFR-1, 3 and VEGFR1 and subsequently inhibition of downstream targets phosphorylated (p)-BAD, p-Foxo1a and p-GSK-3β was observed at 6 hours. NOD/SCID mice implanted with H929 human plasmacytoma xenografts and treated for 30 days with 50, 100, 200mg/kg/d ENMD-2076 showed a dose-dependent inhibition of tumor growth (Figure 1), with minimal toxicity as assessed by the stable weight of treated animals. Immunohistochemical staining of tumors from sacrificed animals showed significant reduction in Ki67 at all dose levels of treatment compared to control tumors. An increase in cleaved caspase-3 was observed on Western blot from the lysates of H929 tumors obtained from treated animals. ENMD-2076 free base also showed synergistic cytotoxic activity when combined with lenalidomide against H929, MM1.R and MM1.S cells as assessed by MTT assay and Annexin-V/PI staining. Using the Chou-Talalay method, the combination indices (CI) were < 1 for all three cell lines across a range of concentrations of ENMD-2076 free base (0.25–1.0 μM) plus lenalidomide (2.5–10 μM) indicating synergistic activity (CI=0.362 H929; CI=0.315 MM1.R; CI=0.415 MM1.S). Our results provide rationale for the investigation of ENMD-2076 alone and in combination with lenalidomide in patients with multiple myeloma. [Display omitted]

Effects of low-dose tamoxifen on breast cancer biomarkers Ki-67, estrogen and progesterone receptors

Breast carcinoma is the most common malignancy among women and it has a major impact on mortality. Studies of primary chemoprevention with tamoxifen have generated high expectations and considerable success rates. The efficacy of lower doses of tamoxifen is similar to that seen with a standard dose of the drug, and there has been a reduction in healthcare costs and side effects.The immune reaction to monoclonal antibody Ki-67 (MIB-1) and the expression of estrogen receptors (1D5) and progesterone receptors (PgR 636) in breast carcinoma were studied in patients treated with 10 mg of tamoxifen for a period of 14 days.A prospective randomized clinical trial was conducted with 38 patients divided into two groups: Group A: N = 20 (control group-without medication) and Group B: N = 18 (tamoxifen/10 mg/day for 14 days). All patients signed an informed consent term previously approved by both institutions. Patients underwent incisional biopsy before treatment and 14 days later a tumor tissue sample was obtained during surgical treatment. Positivity was quantitatively assessed, counting at least 1.000 cells per slide. For statistical data analysis, a Wilcoxon non-parametric test was used, and α was set at 5%.Both groups (A and B) were considered homogeneous regarding control variables. In Group A (control), there was no statistically significant reduction in Ki-67 (MIB-1) (p = 0.627), estrogen receptor (1D5) (p = 0.296) and progesterone receptor positivity (PgR 636) (p = 0.381).In Group B (tamoxifen 10 mg/day), the mean percentage of nuclei stained by Ki-67 (MIB-1) was 24.69% before and 10.43% after tamoxifen treatment. Mean percentage of nuclei stained by estrogen receptor (1D5) was 59.53% before and 25.99% after tamoxifen treatment. Mean percentage of nuclei stained by progesterone receptor (PgR 636), was 59.34 before and 29.59% after tamoxifen treatment. A statistically significant reduction was found with the three markers (p < 0.001).Tamoxifen significantly reduced monoclonal antibody Ki-67 (MIB-1), estrogen receptor (1D5) and progesterone receptor positivity (PgR 636) in the breast epithelium of carcinoma patients treated with a 10 mg dose of tamoxifen for 14 days.