The primary purpose of this study was to evaluate the prognostic significance of Ki-67, a cell proliferation-associated antigen, in a large group (n = 674) of axillary node-negative breast cancer cases with long-term follow-up and to correlate Ki-67 antigen expression with S-phase fraction. Ki-67 immunostaining was assessed both semiquantitatively and quantitatively. The statistical analysis focused on agreement between methods of Ki-67 quantification, agreement between Ki-67 and S-phase fraction, associations between Ki-67 and other clinical variables, and prognostic value of Ki-67. There was excellent agreement between the two methods of Ki-67 assessment (Spearman rank correlation, rsp = 0.91; P = 0.0001; n = 674) but only weak correlation between either semiquantitative or quantitative Ki-67 and S-phase fraction (rsp = 0.12 and rsp = 0.15, respectively). Ki-67 (overall median, 2%) was independent of tumor size and modestly related to other measures of tumor aggressiveness. Using a cutpoint of 5% (percentage of tumor cells), cases with high Ki-67 exhibited a significantly shorter disease-free survival (Padj = 0.004). In multivariate analysis, high Ki-67 was associated with a 1.8-fold increased risk of recurrence (P = 0.001). In the subgroup with S-phase data, the adjusted relative risk (hazard ratio, 1.9; P = 0.02) was unchanged by inclusion of S phase in the model. This suggests that Ki-67 provides significant independent prognostic information in addition to that contained in tumor size and S-phase fraction.
Immunodeficient mice infected with Borrelia turicatae, a relapsing fever agent, have a disorder that resembles disseminated Lyme disease. Two serotypes, A and B, differed in their arthritogenicity in both CB-17 SCID and C3H SCID mice. In CB-17 SCID mice infected with serotype A or B, arthritis was assessed by measurement of tibiotarsal diameter, functional ability on a beam walk test, and microscopic assessment of joint inflammation. Serotype B-infected mice had greater joint swelling, functional disability, and leukocytic infiltration in the joints than serotype A-infected mice. Joint swelling and disability peaked at 2 weeks of infection and then decreased, while leukocyte infiltration in the joints persisted. To investigate the basis for the differences in arthritogenicity of serotypes A and B, spirochete burdens in infected mice were measured by quantitative PCR of spirochete DNA in joints, direct immunofluorescence of spirochetes in joints, and counts of spirochetes in the blood. At 2 weeks of infection there were seven times more spirochetes in the joints of serotype B-infected mice than in those of serotype A-infected mice, measured by both quantitative PCR and direct enumeration. Although serotypes A and B had the same infectivity and growth rate in vivo, serotype B spirochetes were eightfold more abundant in the blood than serotype A spirochetes and produced greater fatality in newborn mice. These findings indicate that differences in disease severity in mice infected with serotype A or B are attributable to differences in the spirochete burden in the joints and blood.
Abstract Background: The Preoperative Endocrine Prognostic Index (PEPI) scores the independent prognostic effects of tumor pathologic staging and expression levels of ER and the “proliferation” marker Ki67 in the surgical sample to predict long term outcomes after completion of neoadjuvant endocrine treatment (Ellis et al JNCI 100:1380, 2008). A limitation of the PEPI is that the prognostic information becomes available only after 4 months of treatment. We therefore evaluated the value of an early assessment of the Ki67 level in a tumor biopsy sample taken two to four weeks after initiating treatment in two neoadjuvant endocrine therapy trials for the purposes of the early identification of non- respondersMethods: A Ki67 cut point of greater than 10% for poor outcome in ER+ breast cancer was derived by comparing the PAM50 intrinsic subtype profile using a qRT-PCR assay with Ki67 data in a 700+ sample data set. A baseline level of 10% or less correlated most closely with a PAM50-based definition of LumA breast cancer and above 10% LumB breast cancer. We subsequently applied the 10% cut point to the baseline and early on-treatment Ki67 data in two trials, POL (Olson et al JACS 208:906, 2009) and IMPACT (Smith et al JCO: 23, 5108, 2005).Results: At baseline the dichotomized Ki67 definition was not significantly predictive for surgical Ki67 level, PEPI score or RFS in this modest size sample set. In contrast, in a result that emphasizes the enhaced prognostic properties of the on-treatment Ki67 approach, the one month POL sample Ki67 values (62 patients) predicted a higher level of Ki67 in the surgical samples at four months after treatment initiation (P=.01), a poorer PEPI score (P=0.01), a smaller number of patients in the PEPI risk point zero group (P=0.08) and worse relapse free survival (P=0.003). The IMPACT data (153 patients) confirmed that a two week Ki67 >10% predicted higher Ki67 in the surgical specimen (P=0.001), a poorer PEPI score (P=0.001), smaller numbers of patients in the PEPI 0 risk point group (P= 0.004) and worse relapse free survival (P=0.008).Ki67 and OutcomePOL 4W Ki67% PEPI 0RFS (events)10%>1/19 (5%)5/21 (23%)10%≤10/36 (28%)1/41 (2.4%)P ValueP=0.08 (Fisher)P=0.003 (log rank)IMPACT 2W Ki67% PEPI 0RFS (events)10%>0/32 (0%)9/35 (26%)10%≤21/101 (21%)13/118 (11%)P ValueP=0.004 (Fisher)P=0.008 (log rank) Conclusions: A tumor Ki67 assessment taken a short time (2 to 4 week window) after the initiation of neoadjuvant AI identifies patients with poor outcome ER+ disease. Amendment 6 of the neoadjuvant endocrine therapy protocol ACOSOG Z1031 will triage patients with an “on treatment” Ki67 value above 10% to chemotherapy in order to assess the pathological response rate to cytotoxic therapy in this important tumor subset.Supported by R01 CA095614, Avon PFP award 3P50 CA68438-07S2, U01 CA114722, ACOSOG U10 CA 76001, Breakthrough Cancer UK and AstraZenica (IMPACT trial). Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 78.
Abstract Background: Neoadjuvant endocrine therapy (NET) trials serve to inform design and selection of agents for larger adjuvant endocrine trials. The neoadjuvant aromatase inhibitor (AI) trial ACOSOG Z1031 randomized the treatment of postmenopausal women with ER rich (Allred 6-8) clinical stage 2/3 breast cancer to either EXE, ANA and LET. The results are therefore of interest in the context of new data that compares these AIs in the adjuvant setting. The opportunity to develop predictive biomarker tests for NET sensitivity was also a key study objective in order guide future selection of patients for inclusion in studies that compare NET versus neoadjuvant chemotherapy (NCT). The value of a baseline PAM50 intrinsic subtype test for NET patient selection was therefore an embedded objective of the study. Methods: Of the 374 women who began study treatment on Z1031, pretreatment biopsy specimen for central IHC testing for ER and Ki67 was available on 304 patients. The PAM50 intrinsic subtype test (PMCID: 2667820) could be conducted on 170 baseline samples. The Preoperative Endocrine Prognostic Index (PEPI) an approach to predicting long term outcomes based on pathological stage and surgical specimen ER and Ki67 status was also conducted because patients with a PEPI of zero (T1/2 N0, ER+, Ki67<=2.7%) have been found to have an excellent long term prognosis and therefore unlikely to benefit from chemotherapy (PMCID: 255670). Results: The degree of Ki67 suppression, the absolute level of Ki67 in the surgical specimen and the PEPI results were not significantly different by treatment arm (Kruskal-Wallis tests p >0.05). Thirty Percent (95%CI: 19.6-42.9%) of women with pre-AI LumA subtype and 12% (95%CI: 6.4-20.0%) of women with pre-AI LumB subtype had a PEPI of zero predicting excellent prognosis. Multivariate logistic modeling found that the likelihood of a PEPI score of zero is 3.2 fold higher (95%CI: 1.43 7.12) among 66 women with a pre-AI LumA subtype compared to 100 women with a pre-AI LumB subtype. Additional baseline variables (clinical T and N stage, HER2 status, pre-AI Ki-67 level (< = 10% vs. >10%) were not found to modify the chance of a PEPI of zero. Baseline PAM50 Subtype versus PEPI Conclusion: The results of Z1031 show no differences in clinical or biologic response between LET, EXE, and ANA in the neoadjuvant setting; predicting that no difference in efficacy will be seen between these agents in adjuvant studies. Pre-AI LumA versus LumB status, defined by the PAM50 subtype, identifies patients who are more likely to be in the most favorable PEPI score following neoadjuvant AI therapy. Patients with pretreatment LumA status may, therefore, not be suitable for a randomized trial of NET versus NCT since approximately one third of these patients may be able to avoid chemotherapy altogether. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr S1-2.
Mutational inactivation of the retinoblastoma gene (RB) is found in all retinoblastomas and in a subset of other human neoplasms, including sarcomas of bone or soft tissue and carcinomas of lung or breast. Exogenous copies of wild-type RB have been shown to suppress the tumorigenicity of several types of tumor cells with endogenous RB mutations, including a previously described human prostatic carcinoma cell line. To further support a role for RB inactivation in the genesis of prostate cancer, seven primary or metastatic prostate carcinoma specimens were examined for evidence of RB mutation. By the use of immunoblot analysis and immunostaining of histologic sections, RB-encoded protein was readily detected in tumor cells of five specimens, was equivocally detected in one specimen, and was apparently absent from tumor cells of one specimen. RB mutations in the latter case were precisely characterized as (i) a deletion of 103 nucleotides containing transcriptional start sites and (ii) loss of the second RB allele. The 103-base-pair deletion was sufficient to abolish the promoter activity of upstream DNA sequences in a heterologous expression system. These results (i) demonstrate that RB can be inactivated in vivo by mutation of its promoter, (ii) confirm the existence of RB mutations in some human prostate carcinomas, and (iii) suggest the use of immunohistochemical methods to screen for RB mutations in clinical samples of common adult neoplasms.
Abstract Background: Neoadjuvant aromatase inhibitor (AI) therapy would become a more acceptable alternative to chemotherapy if there was a way to identify AI-resistant cases early for triage to alternate systemic treatment. We therefore conducted a Phase 2 trial of post-menopausal patients with clinical stage 2 or 3 ER+ (Allred Score 6 to 8) breast cancer who were re-biopsied 2 to 4 weeks after initiating neoadjuvant AI therapy. If the tumor Ki67 level was > 10% (AI resistant) the patient was triaged to either chemotherapy or immediate surgery and if < 10% (AI sensitive) the patient completed 16 to 18 weeks of neoadjuvant AI treatment. Co-primary endpoints were: 1) to confirm that the pathological complete response (pCR) rate to standard NCCN chemotherapy in the AI resistant population is in the range of 20% (similar to ER− disease). This required the observation of least 4/35 pCRs; 2) whether patients were willing to accept a recommendation of no chemotherapy when the preoperative endocrine prognostic index score was zero (PEPI-0) (pStage 1 or 2A, Ki67<2.7% and ER+) since, according to the PEPI model, these patients have such a low relapse risk that chemotherapy is unlikely to be of value (Ellis, M JNCI 100:1380–8, 2008). Feasible was defined as <10% PEPI-0 cases receiving adjuvant chemotherapy. Secondary endpoints included a determination of the distribution of intrinsic subtypes in the AI sensitive and resistant groups (using a research use only assay). Results: 51 patients (21%) had 2 to 4 week Ki67>10%, of these 36 received chemotherapy per protocol (27) anthracycline (A) and taxane (T)-based, 7 T-based, 1 A-only based and 1 other (non-NCCN) regimen but only 2 patients had a pCR (5.5%). 194 patients (79%) had a Ki67 <10%, of these 176 had PEPI score information. 65 had a PEPI-0 (37%), of which 61 (94%) reported a treatment plan that did not include adjuvant chemotherapy. The AI resistant group had a significantly higher frequency of baseline high-risk intrinsic subtypes (LumB: 26, HER2-E: 2 versus LumA: 7) at baseline than the AI sensitive group (LumB: 40, HER2-E: 0 versus LumA: 66) Fishers exact test p = 7.16E−6. Additional molecular data will be presented at the meeting. Conclusion. A Ki67 value >10% at 2 to 4 weeks strongly enriches for high-risk molecular subtypes (mainly LumB) however standard neoadjuvant chemotherapy did not meet pre-assigned criteria for adequate Phase 2 activity in this group. Novel neoadjuvant approaches for AI resistant-disease defined by high on treatment Ki67 are therefore a high priority. Management of patients with excellent response to neoadjuvant endocrine therapy (PEPI-0) without adjuvant chemotherapy is feasible. The ALTERNATE trial will enroll over 2000 patients to establish a new standard of care for neoadjuvant treatment of ER+ HER2− disease based on these principles. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr PD07-01.
Abstract Purpose This goal of this study was to determine the impact of vascular motion on acute drug transfer and retention of drug-coated balloons (DCB) or drug-eluting stents (DES). Methods Commercially available paclitaxel DCBs (Lutonix & IN.PACT) and a paclitaxel DES (Zilver) were subjected to physiological flow and vascular motion conditions using a peripheral-simulating benchtop bioreactor system. Each DCB- or DES-treated artery was subjected to three sets of movement parameters including pulsatile flow with no twisting/bending (P1), pulsatile flow with 16.8° twist, 25° bend and 3.2 mm compression (P2), and pulsatile flow with 68° twist, 35° bend, 21 mm compression (P3). After 24 h, the treated segments were removed and paclitaxel concentrations were measured using pharmacokinetic analysis. Results In the group of arteries treated with the Lutonix DCB, there was a significant decrease in arterial paclitaxel concentrations between the P1 and both the P2 and P3 moving parameters (P1 = 404 ± 195 ng/mg, P2 = 14.9 ± 9.92 ng/mg, P3 = 19.2 ± 15.4 ng/mg; P1-P2 p = 0.007, P1-P3 p = 0.005). For the IN.PACT DCB group, no differences in the mean arterial paclitaxel concentrations were observed for the various movements ( p = 0.55). Lastly, in the Zilver DES group, differences were only measured between the P2 and P3 moving parameters (P2 = 84.8 ± 32.7 ng/mg, P3 = 0.11 ± 0.06 ng/mg; P2-P3 p = 0.01). Conclusion Acute retention of arterial paclitaxel levels can be adversely impacted by vascular movement in both DES- and DCB- treated arteries.
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