The role of infiltrating lymphocytes in the neo-adjuvant treatment of women with HER2-positive breast cancer.

BACKGROUND: Pre-treatment tumour-associated lymphocytes (TILs) and stromal lymphocytes (SLs) are independent predictive markers of future pathological complete response (pCR) in HER2-positive breast cancer. Whilst studies have correlated baseline lymphocyte levels with subsequent pCR, few have studied the impact of neoadjuvant therapy on the immune environment. METHODS: We performed TIL analysis and T-cell analysis by IHC on the pretreatment and 'On-treatment' samples from patients recruited on the Phase-II TCHL (NCT01485926) clinical trial. Data were analysed using the Wilcoxon signed-rank test and the Spearman rank correlation. RESULTS: In our sample cohort (n = 66), patients who achieved a pCR at surgery, post-chemotherapy, had significantly higher counts of TILs (p = 0.05) but not SLs (p = 0.08) in their pre-treatment tumour samples. Patients who achieved a subsequent pCR after completing neo-adjuvant chemotherapy had significantly higher SLs (p = 9.09 × 10-3) but not TILs (p = 0.1) in their 'On-treatment' tumour biopsies. In a small cohort of samples (n = 16), infiltrating lymphocyte counts increased after 1 cycle of neo-adjuvant chemotherapy only in those tumours of patients who did not achieve a subsequent pCR. Finally, reduced CD3 + (p = 0.04, rho = 0.60) and CD4 + (p = 0.01, rho = 0.72) T-cell counts in 'On-treatment' biopsies were associated with decreased residual tumour content post-1 cycle of treatment; the latter being significantly associated with increased likelihood of subsequent pCR (p < 0.01). CONCLUSIONS: The immune system may be 'primed' prior to neoadjuvant treatment in those patients who subsequently achieve a pCR. In those patients who achieve a pCR, their immune response may return to baseline after only 1 cycle of treatment. However, in those who did not achieve a pCR, neo-adjuvant treatment may stimulate lymphocyte influx into the tumour.

Expression of anaesthetic and analgesic drug target genes in excised breast tumour tissue: Association with clinical disease recurrence or metastasis.

BACKGROUND: Retrospective analyses suggest anaesthetic-analgesics technique during cancer surgery may affect recurrence/metastasis. This could involve direct effects of anaesthetic-analgesic drugs on cancer cells. While µ-opioid receptor over-expression in lung tumours is associated with greater metastasis, other anaesthetic-analgesic receptor targets in cancer recurrence/metastasis remain unexplored. Therefore, we evaluated the association between genetic expression of anaesthetic-analgesic receptor targets and recurrence/metastasis, using a repository of breast cancer gene expression and matching clinical data. METHODS: A list of 23 genes encoding for the most prominent anaesthetic-analgesic receptor targets was compiled. This was processed through BreastMark- an algorithm integrating gene expression data from ~17,000 samples and clinical data from >4,500 breast cancer samples. Gene expression data was dichotomized using disease-free survival (survival without recurrence) and distant disease-free survival (survival without metastasis) as end points. Hazard ratios were calculated by Cox-regression analysis. Enrichment for prognostic markers was determined by randomly choosing 23-member gene lists from all available genes, calculating how often >5 significant markers were observed and adjusting p-values for multiple testing. This was repeated 10,000 times and an empirical p-value calculated. RESULTS: Of 23 selected genes, 9 were significantly associated with altered rates of metastasis and 4 with recurrence on univariate analysis. Adjusting for multiple testing, 5 of these 9 genes remained significantly associated with metastasis, non with recurrence. This ratio of genes (5/23) was not significantly enriched for markers of metastasis (p = 0.07). CONCLUSION: Several anaesthetic-analgesic receptor genes were associated with metastatic spread in breast cancer. Overall there was no significant enrichment in prognostic markers of metastasis, although a trend was observed.

The prognostic utility of the transcription factor SRF in docetaxel-resistant prostate cancer: in-vitro discovery and in-vivo validation.

BACKGROUND: Docetaxel based therapy is one of the first line chemotherapeutic agents for the treatment of metastatic castrate-resistant prostate cancer. However, one of the major obstacles in the treatment of these patients is docetaxel-resistance. Defining the mechanisms of resistance so as to inform subsequent treatment options and combinations represents a challenge for clinicians and scientists. Previous work by our group has shown complex changes in pro and anti-apoptotic proteins in the development of resistance to docetaxel. Targeting these changes individually does not significantly impact on the resistant phenotype but understanding the central signalling pathways and transcription factors (TFs) which control these could represent a more appropriate therapeutic targeting approach. METHODS: Using a number of docetaxel-resistant sublines of PC-3 cells, we have undertaken a transcriptomic analysis by expression microarray using the Affymetrix Human Gene 1.0 ST Array and in conjunction with bioinformatic analyses undertook to predict dysregulated TFs in docetaxel resistant prostate cancer. The clinical significance of this prediction was ascertained by performing immunohistochemical (IHC) analysis of an identified TF (SRF) in the metastatic sites from men who died of advanced CRPC. Investigation of the functional role of SRF was examined by manipulating SRF using SiRNA in a docetaxel-resistant PC-3 cell line model. RESULTS: The transcription factors identified include serum response factor (SRF), nuclear factor kappa-B (NFκB), heat shock factor protein 1 (HSF1), testicular receptor 2 & 4 (TR2 &4), vitamin-D and retinoid x receptor (VDR-RXR) and oestrogen-receptor 1 (ESR1), which are predicted to be responsible for the differential gene expression observed in docetaxel-resistance. IHC analysis to quantify nuclear expression of the identified TF SRF correlates with both survival from date of bone metastasis (p = 0.003), survival from androgen independence (p = 0.00002), and overall survival from prostate cancer (p = 0.0044). Functional knockdown of SRF by siRNA demonstrated a reversal of apoptotic resistance to docetaxel treatment in the docetaxel-resistant PC-3 cell line model. CONCLUSIONS: Our results suggest that SRF could aid in treatment stratification of prostate cancer, and may also represent a therapeutic target in the treatment of men afflicted with advanced prostate cancer.

ADAM-17: a novel therapeutic target for triple negative breast cancer.

BACKGROUND: Validated targeted therapy is currently unavailable for patients with invasive breast cancer negative for oestrogen receptors, progesterone receptors and HER2 [i.e., those with triple-negative (TN) disease]. ADAM-17 is a protease involved in the activations of several ligands that bind to and promotes intracellular signalling from the EGFR/HER family of receptors. PATIENTS AND METHODS: Expression of ADAM-17 was measured in 86 triple-negative and 96 non-triple-negative breast cancers. The ADAM-17 specific inhibitor, PF-5480090 (TMI-002, Pfizer) was tested in a panel of breast cancer cell lines for effects on functional outputs. RESULTS: In this study we show using both Western blotting and immunohistochemistry that ADAM-17 is expressed at significantly higher levels in TN than non-TN breast cancers. Using a panel of breast cancer cell lines in culture, PF-5480090 was found to decrease release of the EGFR ligand, TGF-alpha, decrease levels of phosphorylated EGFR and block cell proliferation in a cell-type-dependent manner. Potentially important was the finding of a significant and moderately strong correlation between ADAM-17 activity and extent of proliferation inhibition by PF-5480090 (r = 0.809; p = 0.003; n = 11). Pretreatment of cell lines with PF-5480090 enhanced response to several different cytotoxic and anti-EGFR/HER agents. CONCLUSION: It is concluded that inhibition of ADAM-17, especially in combination with chemotherapy or anti-EGFR/HER inhibitors, may be a new approach for treating breast cancer, including patients with TN disease.

Comprehensive/tertiary care for headache: a 6-month outcome study.

OBJECTIVE: To assess programwide (outpatient plus inpatient) outcome using prospective measures for the first 6 months of treatment at a comprehensive headache center. Background.-There is little published data on the overall programwide efficacy of comprehensive, multidisciplinary treatment centers for severe, refractory headache disorders. METHODS: For 1 week each month over a 2-year period, all new patients completed an initial questionnaire noting frequency/severity of headaches and other headache-relevant measures. A follow-up version mailed at 6 months was returned by 218 of 421 patients (response rate 52%). Sixteen percent were treated both in and out of the hospital, with 84% treated as outpatients only. Patients had a mean of 2.50 medical visits after the initial evaluation, and 43% met with a psychologist. RESULTS: Mean headache frequency dropped from 5.30 to 3.45 days per week (P<.00001), with severe headaches declining from 2.31 to 1.33 days per week (P<.00001). Sixty-seven percent of the patients had at least a 50% reduction in headaches, with a mean percentage improvement per patient in severe headaches of 56% (median 67%). Emergent care visits within 6 months dropped from 4.39 to 1.67 (P<.00001). Days with significant headache-related work impairment declined from 2.04 to 0.67 days per week (P<.00001), a net annualized reduction of 71.24 impaired workdays per year per patient. For full-time workers, missed work days in 6 months dropped from 5.46 to 2.68 (P<.00261). The mean percentage improvement for headache-impaired workdays was 67% (median 88%), and for work absence was 63% (median 100%). Total days incapacitated per week dropped from 1.72 to 0.89 (P<.00001), with a mean percentage improvement of 67% (median 91%). Significant treatment satisfaction was reported by 89%. A weighted Goal Attainment Scale based on the percentage of patients showing significant improvement in pain control, functioning, work performance, reliance on emergent care, depression, and satisfaction revealed an overall Goal Attainment score of 72%. CONCLUSIONS: Despite a mean of only 3.50 medical visits in a population of complex patients, significant improvement was demonstrated in several key economically relevant variables within 6 months of referral. If maintained over the long term, the results support the concept that matching intensity of treatment (comprehensive/tertiary care) to severity of illness (complex/refractory cases) is cost-effective.

Comprehensive inpatient treatment for intractable migraine: a prospective long-term outcome study.

This paper describes outcome data for 100 patients with severe, intractable, persistent migraine (chronic daily headache), who were admitted to a comprehensive inpatient Head-Pain Treatment Unit for a mean of 8.5 days. Headache, pain-related behavior, depression, sleep disturbance, functional performance, work status, and medication use were assessed at admission, post-discharge (mean = 2.3 weeks) and long-term follow-up (mean = 8.3 months). Significant improvement noted two weeks after discharge was maintained over time. Long-term results revealed a 64% reduction in the mean number of days in a 2-week period with severe to incapacitating headache (6.29 to 2.26), with a corresponding increase in pain-free days (1.03 to 5.40). At follow-up, the frequency of severe headaches was reduced by at least 50% for 75% of the patients. The mean rating of overall improvement was 74%. Patients on work-leave due to pain dropped from 24% to 4%, while the number of working patients rose from 31% to 53%. The data showed statistically significant reductions in days lost to pain, depression, sleep disturbance, and use of symptomatic medication. All the above analyses were significant at P = .000. These results demonstrate the efficacy of the inpatient headache program intervention for this group of patients.