Our cluster analysis of the Cancer Genome Atlas for co-expression of HSP27 and CRYAB in breast cancer patients identified three patient groups based on their expression level combination (high HSP27 + low CRYAB; low HSP27 + high CRYAB; similar HSP27 + CRYAB). Our analyses also suggest that there is a statistically significant inverse relationship between HSP27 and CRYAB and known clinicopathological markers in breast cancer. Screening an unbiased 248 breast cancer patient tissue microarray (TMA) for the protein expression of HSP27 and phosphorylated HSP27 (HSP27-82pS) with CRYAB also identified three patient groups based on HSP27 and CRYAB expression levels. TMA24 also had recorded clinical-pathological parameters, such as ER and PR receptor status, patient survival, and TP53 mutation status. High HSP27 protein levels were significant with ER and PR expression. HSP27-82pS associated with the best patient survival (Log Rank test). High CRYAB expression in combination with wild-type TP53 was significant for patient survival, but a different patient outcome was observed when mutant TP53 was combined with high CRYAB expression. Our data suggest that HSP27 and CRYAB have different epichaperome influences in breast cancer, but more importantly evidence the value of a cluster analysis that considers their coexpression. Our approach can deliver convergence for archival datasets as well as those from recent treatment and patient cohorts and can align HSP27 and CRYAB expression to important clinical-pathological features of breast cancer.

Abstract. Early detection of lung cancer may potentially help to improve the outcome of this fatal disease. Currently, no satisfactory laboratory tests are available to screen for this type of cancer. The aim of this study was to improve diagnostic procedures for lung cancer through the discovery of serum biomarkers using SELDI-TOF MS (surfaceenhanced laser desorption/ionization time-of-flight mass spectrometry). Mass spectrometric profiling was applied to the serum of a total of 139 lung cancer patients and 158 healthy individuals for developing a prognostic signature. For validation, two separate groups were employed, comprising of 126 and 50 individuals, respectively. Informative regions of mass spectra were identified by forming protein mass clusters and identifying predictive clusters in a logistic regression model. A total of 17 differential predictive protein mass clusters were identified in patients with metastatic lung cancer disease compared to healthy individuals. These clusters provide for a robust risk prediction model. The sensitivity and specificity of this model was estimated to be 87.3 and 81.9%, respectively, for the first validation set, and 96.0 and 66.7%, respectively, for a second validation set of patients with early disease (stages I and II). A differential 11.5/11.7 kDa double-peak could be identified as serum amyloid · (SAA) by peptide mapping. Our data suggest that the SELDI-TOF MS technology in combination with a careful statistical analysis appears to be a useful experimental platform which delivers a rapid insight into the proteome of body fluids and may guide to identify novel biomarkers for lung cancer disease.

Background: Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) is a protein complex found in almost all animal cell types and is involved in cellular responses to stimuli such as stress, free radicals, cytokines et.c. NF-kB plays a key role in regulating the immune response and incorrect regulation of NF-kB has been linked to the development of cancer, several inflammatory and autoimmune diseases. The role of NF-kB, its subunits and pathways in breast cancer and in the surrounding stromal tissues remains uncertain. αB-crystallin is a heat shock protein, which function as stress-induced molecular chaperones to inhibit the aggregation of denatured proteins. Previous studies have identified αB-crystallin as a marker of poor prognosis for breast cancer and have suggested that it is an excellent marker for tumours of basal origin. The aim of this study is to examine, for the first time, NF-kB and αB-crystallin, vimentin and HSPs in the same set of breast cancers and stroma and relate this to clinical parameters including outcome.Methods: Tissue Micro Arrays of 0.6mm cores of 246 breast cancers were stained with antibodies to αB-crystallin, vimentin, HSP27 (antibody ERD5) and HSP27 82P and 12 subsequent antibodies for NF-kB (IKKa, Ser276, Ser32/36, 180/181, P52, 1kkygmma, P65, RELB, P100/P52, IkBa, cREL, and P50) and scored using the Quick Score Method.Results: Positive associations have been found between αB-crystallin and IKBA (P=0.001 Fishers exact test (FET)), RELB (P<0.001 (FET)) and SER32/36 (P=0.03 (FET)) whereas Vimentin was positively associated IKBA (P=0.003 (FET)), IKKA (P=0.001 (FET)), P50 (P=0.002 (FET)), RELB (P<0.001 (FET)) and SER32/36 (P=0.01 (FET)). Positive associations have also been found between HSP27 and P50 (P=0.002 (FET)), SER32/36 (P<0.001 (FET)). Antibodies such as 180/181 (P=0.04 Kaplan Meier Log Rank (KM), cREL (P=0.03 (KM)), IKBA (P=0.03 (KM)) and IKKA (P=0.01 (KM)) was associated with good survival and P50 (P=0.002 (KM)) and P65 (P=0.01 (KM)) to poor survival. Expression of αB-crystallin was associated with vimentin (P<0.001 (FET)). Its expression was linked to a low expression of the estrogen receptor. Vimentin expression was associated with estrogen receptor (ER) negative cancers and poor survival (P <0.001 (FET)), P=0.002 (KM Log Rank) respectively). In contrast to αB-crystallin, low expression of HSP27 was associated with low ER and progesterone receptor (PGR).Conclusions: Associations were found between various antibodies within the NF-kB pathway as well as the heat shock proteins αB-crystallin, vimentin and HSP27 with varying implications for overall survival. We aim to clarify the mechanisms to prevent the aggregation of stress-accumulated misfolded proteins and apoptosis resulting better survival. Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 2143.

#5070 Background: Previous studies have identified αB-crystallin as a marker of poor prognosis for breast cancer and have suggested that it is an excellent marker for tumours of basal origin. Increased expression of αB-crystallin has been associated with anchorage-independent growth and metastasis leading to the suggestion that αB-crystallin is an oncoprotein. We have considered whether the αB-crystallin binding proteins, vimentin and HSP27 also show a similar association with poor prognosis. We also investigated whether either HSP27 phosphorylation HSP27 or p53 expression could be associated with αB-crystallin expression.
 Methods: Tissue Micro Arrays of 0.6x0.6mm cores of 205 breast cancers were stained with antibodies to αB-crystallin, vimentin, p53 (antibodies DO1, FP3), HSP27 (antibody ERD5) and HSP27 82P. The levels of protein expressions were then compared with clinical and pathological parameters.
 Results: The expression of αB-crystallin was positively associated with vimentin (P<0.001 Fisher's exact test (FET)). We have confirmed that αB-crystallin expression is linked to a low expression of the estrogen receptor and reduced survival (P<0.001 (FET), P=0.002 Kaplan Meier Log Rank (KM) respectively). Vimentin expression was similarly associated with estrogen receptor (ER) negative cancers and poor survival (P <0.001 (FET), P= 0.002 (KM Log Rank) respectively). Low expression of HSP27 resulted in poor three-year survival (P=0.007 (KM Log Rank)), but longer-term survival was unaffected (P=0.09 (KM Log Rank)). In contrast to αB-crystallin, low expression of HSP27 was associated with ER and progesterone receptor (PGR) negativity (P<0.001 (FET), P=0.001 (FET)). HSP27 82P similarly resulted in poor three-year survival (P=0.01 (KM Log Rank)) compared with longer-term survival (P=0.05 (KM Log Rank)). In samples positive for either αB-crystallin or vimentin, a strong association with increased p53 expression (p53 antibodies: DO1(P<0.001 (FET)) and FP3(P<0.001 (FET))) was found.
 Conclusions: The increased expression of the protein chaperone, αB-crystallin was linked with reduced survival as well as it's binding partner, vimentin. We found that within this patient subgroup, there was an increased level of p53. The potential functional significance of this interaction for metastasis will be discussed in the context of other predictive markers for breast cancer. A similar association was not found for HSP27 expression. Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 5070.