<p> Women who undergo immediate mastectomy for breast cancer experience tremendous anxiety when faced with breast reconstruction and are often conflicted regarding which type of breast reconstruction to choose. This pilot study aimed to analyze the impact of a decision aid, adapted from Stanford University Breast Center, on decisional conflict in women with newly diagnosed breast cancer seeking breast reconstruction. Twenty newly-diagnosed breast cancer patients seeking breast reconstruction at a large academic healthcare center were randomized into two groups. Comparisons were made between women who reviewed the standard educational materials prior to initial consultation, and women who reviewed these materials and then reviewed a decision aid brochure at initial consultation and two weeks post consultation, utilizing the Decisional Conflict Scale. Technical issues halted data collection after only nine participants completed the study. Although no reliable findings could be interpreted from such a small sample size, the results suggest the decision aid as a valuable tool for patient education. Nurses may gain increased awareness of the emotional conflicts faced by newly diagnosed breast cancer patients when making decisions for breast reconstruction.</p><p>

No description available.

abstract: CpG methylation is an essential requirement for the normal development of mammals, but aberrant changes in the methylation can lead to tumor progression and cancer. An in-depth understanding of this phenomenon can provide insights into the mechanism of gene repression. We present a study comparing methylated DNA and normal DNA wrt its persistence length and contour length. Although, previous experiments and studies show no difference between the physical properties of the two, the data collected and interpreted here gives a different picture to the methylation phenomena and its effect on gene silencing. The study was extended to the artificially reconstituted chromatin and its interactions with the methyl CpG binding proteins were also probed. / Dissertation/Thesis / Ph.D. Physics 2012

<p> Pancreatic ductal adenocarcinoma (PDAC) is a notoriously lethal disease characterized by prominent stromal involvement, which plays complex roles in regulating tumor growth and therapeutic response. The extracellular matrix (ECM)-rich stroma has been implicated as a barrier to drug penetration, although stromal depletion strategies have had mixed clinical success. It remains less clear how biophysical interactions with the ECM regulate invasive progression and susceptibilities to specific therapies. Here, an integrative approach combining 3D cell culture and quantitative imaging techniques is used to evaluate invasive behavior and motility as determinants of response to classical chemotherapy and photodynamic therapy (PDT), in which light activated agents induce site-directed cell death by generating reactive oxygen species. The 3D culture protocol developed for these studies with transplanted multicellular PDAC spheroids in rheologically characterized ECM shows that in invasion-promoting ECM environments, PDT response is markedly enhanced in the most motile populations while the same cells exhibit chemoresistance. Conversely, drug-resistant sublines with characterized increase in invasive potential were generated to compare differential treatment response in identical ECM conditions, monitored by particle-tracking microrheology measurements of matrix remodeling. In both scenarios, ECM infiltrating cells exhibit increased PDT sensitivity, whether invasion is consequent to selection of chemoresistance, or whether chemoresistance is correlated with acquisition of invasive behavior. However, while ECM-infiltrating, chemoresistant cells exhibit mesenchymal phenotype, EMT induction in monolayers lacking ECM is not sufficient to enhance PDT sensitivity, yet does impart chemoresistance as expected. In further experiments seeking to elucidate intertwined roles of mechanical and biochemical interactions with ECM components, invasive progression and response to therapeutics were evaluated using ECM protein admixtures and collagen hydrogels with varying extent of crosslinking. In these studies, increased collagen stiffness or presence of laminin-rich ECM both inhibit invasion of PDAC cells, although cells that do infiltrate into ECM nevertheless exhibit chemoresistance and enhanced PDT sensitivity, independent of their ECM environment. In addition to containing platform development with broader applicability to inform microenvironment-dependent therapeutics, results of this work collectively reveal the efficacy of PDT for targeting the most aggressive, chemoresistant, and invasive PDAC cell populations associated with dismal outcomes for this disease.</p><p>

Squamous cell carcinoma (SCC) is the most common histological type of cancer in the head and neck region and arises in the epithelial mucosa of the upper aerodigestive tract. Approximately one and a half million people are living with the diagnosis. Despite efforts in prevention and advances in treatment, the 5-year survival rate still lies around 60%, and recurrences and second primary tumors remain a problem. Moreover, treatment responses vary from patient to patient, highlighting the need for individually tailored treatments. To make this possible, biomarkers predicting treatment outcome are needed to better guide treatment decisions. The aim of this thesis was to evaluate the expression of certain proteins and the frequency of certain SNPs (Single nucleotide polymorphisms) in tumor biopsies and cell cultures of head and neck squamous cell carcinomas (HNSCC), and to explore their potential as biomarkers for treatment outcome. Furthermore, we aimed to study the impact of hypoxia on treatment response, epithelial-tomesenchymal transition (EMT), and induction of cancer stem cells (CSC). In papers I and II, we investigated two proteins, survivin and WRAP53β, using immunohistochemistry (IHC) in tumor biopsies from 40 patients categorized as Non-responders or Responders to radiotherapy. High expression of survivin and nuclear expression of WRAP53β were significantly more prevalent in the Responder group. The combination of these two factors correlated strongest to overall survival, but not to a significantly higher extent compared to survivin alone. Moreover, when examined separately, a high percentage of p53-stained cells and the presence of the SNP FGFR4 Gln388Arg correlated to improved overall survival, whereas the SNP XPD Lys751Gln was associated with worse overall survival. The latter three showed no significant correlations to radiotherapy response. In paper III, the two most promising proteins identified in papers I and II were analyzed in a study cohort of 149 tumor biopsies of glottic laryngeal SCC, categorized as T2N0-T3N0. In this patient group, no significant associations between survivin expression and survival could be found. However, expression of cytoplasmic WRAP53β was significantly linked to worse disease-free-survival (DSF) compared to nuclear WRAP53β or negative staining for WRAP53β. Positive expression of p16INK4a was found in 7% of the tumors. The prevalence of p16 INK4a was higher in younger patients (&lt;60) and associated with absence of recurrence and longer DSF. In paper IV, five HNSCC cell lines were cultured in normoxic (20% O2) and hypoxic (1% O2) conditions and changes in treatment response, EMT profile, and expression of CSC markers were examined. As expected, hypoxia induced EMT and to a certain extent expression of CSC markers. Silencing of the hypoxia-inducible-factor-1α (HIF-1α) only partly reversed these effects, suggesting that other mechanisms are involved. Whereas most cell lines became more resistant to treatment in hypoxia, one cell line (LK0412) became more sensitive to cetuximab-treatment in hypoxia, an effect that was revoked by depletion of HIF-1α, suggesting a possible sensitizing effect of HIF-1α to cetuximab-treatment. Taken together, WRAP53β appears to be a promising biomarker candidate for treatment outcome in HNSCC, but further evaluation especially on the subcellular localization of WRAP53β is required. Even though the role of survivin in radiotherapy response in glottic SCC seems to be insignificant, it might have a more important role in other HNSCC subsites. As far as the effects of hypoxia, it appears that hypoxia might have a sensitizing effect on cetuximab-treatment in certain cases, which seems to be HIF1-α –dependent. Further studies are required to clarify the importance of this observation.

Oncolytic virotherapy (OV) is an innovative alternative to conventional cancer therapies based on the concept of selecting or engineering viruses to preferentially replicate in and kill tumor cells by exploiting their genetic defects. Intra-tumoural innate immunity plays a significant role in blocking the effective therapeutic spread of OV. Histone deacetylase inhibitors (HDIs) are known to blunt cellular anti-viral response. This research demonstrates that HDIs enhance the sensitivity of various cancer cell lines to the replication and spread of three different OV platforms---vesicular stomatitis virus (VSV), semliki forest virus (SFV) and vaccinia. The increased oncolytic activity of VSV correlated with a dampening of cellular interferon responses and augmentation of virus replication. Enhancement of virus replication post HDI treatment was also observed in multiple primary human tumor explants as well as in tumor bearing in vivo models. These results illustrate the general utility of HDIs as chemical switches to regulate cellular innate antiviral responses and to provide controlled growth of therapeutic viruses within malignancies.

Background. Colorectal cancer is the second leading cause of death from cancer in Canada. Many studies have examined associations between GSTM1, GSTT1, and GSTP1 polymorphisms and colorectal cancer but with conflicting results. The objective of this study was to examine the totality of evidence for association between these polymorphisms and risk for colorectal cancer or adenoma. Methods. A systematic review, meta- and pooled analysis were conducted. Results. The meta- and pooled analysis suggest weak significant associations between GSTM1 (ORpooled=1.11; 95%CI: 1.02,1.23), GSTT1 (ORpooled=1.22; 95%CI: 1.10,1.35), and with presence of null variants of each and colorectal cancer, but with heterogeneity between studies. No association between GSTP1 and colorectal cancer, or between any of the polymorphisms and colorectal adenoma, was observed. Significant multiplicative GST gene-gene interactions or gene-environment (smoking) interaction effects were not observed. Conclusion. Evidence to date shows weak associations between GSTM1 and GSTT1 and colorectal cancer.

<p> Anal squamous cell carcinoma (SCC) which is strongly associated with human papilloma virus (HPV) infection is a rare cancer but its incidence is increasing throughout the world. Even though it represents just 0.4% of all new cancer cases in the US, the mortality rate is estimated at 14%, which is comparable to both breast and prostate cancer mortality rates. To decrease the high rate of mortality and morbidity of anal cancer there is an enormous need for early detection and prevention strategies. Besides understanding the role of HPV infection, we also need to comprehend the basics of genetics and epigenetics involved in anal cancer progression. With both the highest incidence rate and a lower survival rate among African-American men, we are interested in understanding the relationship of HPV, miRNAs and somatic mutations associated with the African-American population in anal cancers. This was accomplished by (1) identifying and determining HPV genotypes associated with anal condylomas, pre-malignant/dysplastic lesions and malignant anal SCC through type specific genotyping, (2) profiling miRNAs in anal SCC based on gender and type of HPV infection to identify novel biomarkers using Nanostring technology, and (3) by identifying oncogenic mutations associated with anal lesions, transformation and progression using novel next generation sequencing methods. Common HPV genotypes associated with our samples included HPV-11, 16, 6, 32, 35, 51, 58, 59, and 68, of which HPV-32, 51, 59 and 68 are not protected by the current FDA approved nonavalent vaccine. Furthermore, 10 of 800 known human miRNAs were significantly dysregulated in SCC samples; these miRNAs (miR-451a, miR-1185-13p, miR-637, miR-4525a-5p, miR-1275, miR-1303, miR-600, miR-892b, miR-297 and miR-944) target tumor suppressor and oncogenes and potentially play an oncomir role in cancer progression. <i>TP53, PIK3CA, PDGFRA, HRAS</i>, and <i> RET</i> were some of the most frequently found somatic mutations in the sample set and it was observed that the accumulation of mutations begin at the condyloma stage. In conclusion, it was determined that three key factors determine the possible progression of anal cancer and can therefore aid in future development of novel targeted therapy approaches: type of HPV infection, epigenetic factors involving miRNAs, and genetic factors such as &lsquo;driver&rsquo; somatic mutations that an individual accumulates over their lifetime.</p><p>

<p> Telomere dysfunction describes the catastrophic damage at telomeres, which often leads to genomic instability at the cellular level. There is rising evidence showing that telomere dysfunction also influences the extracellular environment with the inflammatory response. However, little is known about the molecular mechanism of this dysfunctional telomere-associated inflammation. In this dissertation, we identified extracellular forms of Telomeric repeat-containing RNA (TERRA), and demonstrated it might play a role in mediating the crosstalk of telomere dysfunction and inflammation. We found this cell-free TERRA (cfTERRA) is present in mouse tumor and embryonic brain tissue, as well as in human tissue culture cell lines using RNA in situ hybridization. RNA-seq analyses revealed TERRA to be among the most highly represented transcripts in extracellular fractions derived from both normal and cancer patient blood plasma. By characterizing extracellular fractions of the human lymphoblastoid cell line (LCL) culture media, cfTERRA is shown as a shorter form (&sim;200 nt) of cellular TERRA and co-purifies with CD63- and CD81-positive exosome vesicles that could be visualized by cryo-electron microscopy. Mass spectrometry and extracellular chromatin immunoprecipitation (ChIP) assays revealed that regular cfTERRA was physically interacting with histones and telomeric DNA. Incubation of cfTERRA-containing exosomes with peripheral blood mononuclear cells (PBMCs) stimulated transcription of several inflammatory cytokine genes, including TNF&alpha;, IL6, and C-X-C chemokine 10 (CXCL10). Exosomes engineered with elevated TERRA or liposomes with synthetic TERRA further stimulated inflammatory cytokines, suggesting that exosome-associated TERRA augments innate immune signaling. The levels of cfTERRA and DNA damage marker &gamma;H2AX were increasingly incorporated into the exosomes during telomere dysfunction. These dysfunctional telomere-derived exosomes activated a more robust transcription of inflammatory cytokines in PBMCs. These findings imply a previously unknown extrinsic function of TERRA and a potentially molecular mechanism of communication between telomeres and innate immune signaling in tissue and tumor microenvironments. </p><p>

<p> <b><u>Lenalidomide (CC-5013; REVLIMID, Celgene Corp., NJ, USA) belongs to a new immunomodulatory class of drugs called IMiDs.</u></b> It is an oral thalidomide analogue drug that belongs to the second generation of IMiDs. Its parent compound thalidomide&rsquo;s initial descriptions of its teratogenicity are attributed to its anti-angiogenic properties. The drug has other mechanisms of action beyond just cytotoxicity and immune modulation. It includes effects on both the tumor and the tumor microenvironment. </p><p> Within <b><u>hematological malignancies</u></b>, lenalidomide is being used in a number of conditions. Lenalidomide is being used to treat myelodysplastic syndrome patients with 5q deletion resulting in improvement in their transfusion requirements. Lenalidomide in combination with steroids is used in patients with multiple myeloma. </p><p> Recently there has been increased interest to use lenalidomide to treat patients due to its immunomodulatory effect. According to the Surveillance, Epidemiology, and End Results Program (SEER), Non-Hodgkin Lymphoma (NHL) represents 4.3% of all new patients with cancer in United States, with an estimated 71,850 cases in 2015. Within NHL, diffuse large B-cell lymphoma (DLBCL) is most frequently seen constituting approximately 40% of these cases. </p><p> Therapies upfront (at the time of diagnosis) for DLBCL include chemotherapeutic options in combination with biologics/monoclonal antibodies (mAbs), which are curative in intent. In the <u>relapsed or refractory settings </u>, the intent treatment for patients with DLBCL is to achieve cure; however, these have to be consolidated with a stem cell transplant (autologous in most instances). The number of treatment options is increasing, and newer regimens and classes of drugs are being developed and tested for patients with DLBCL. These go beyond mAbs and traditional chemotherapeutic agents, and include novel targeted therapies, immunotherapies and immunomodulatory agents. Immune modulation, as noted, has been a focus of increasing interest particularly for patients with DBLCL. With lenalidomide, responses were seen in up to half of the patients treated with lenalidomide, with about a quarter achieving complete responses. Even as single agent, these responses appeared to be relatively durable (PFS 6.2 months), given the highly aggressive nature of disease. The side effect profile was noted to be manageable without any untoward adverse events. </p><p> This led to the development of clinical trials for patients incorporating lenalidomide. These were both in frontline as well as the relapsed/refractory setting. Lenalidomide (Revlimid) was added to the frontline regimen R-CHOP [referred to as R2CHOP] and showed significant activity and manageable safety profile. The focus of my thesis is the <b><u>development of the clinical trial of incorporating lenalidomide (Revlimid) in the relapsed/refractory setting</u></b> to one of the most commonly employed chemoimmunotherapy regimens called the R-ICE (rituximab-ifosfamide-carboplatin-etoposide) regimen. The regimen developed therefore, is called R2ICE. </p><p> The patients that we studied, who have refractory disease or relapse with lymphoma, constitute at least a quarter of all patients with DLBCL. The majority of these are patients who relapse within the first 1.5 years of upfront curative treatment. For patients with first-relapse/primary-refractory DLBCL, the response rate achieved prior to proceeding with a stem cell transplant (SCT) is a key variable. Usually this is an autologous stem cell transplant (ASCT). ASCT can be potentially curative for these patients who tend to show chemosensitivity by achieving either a complete response (CR) or partial response (PR) with their salvage chemotherapy prior to the transplant. Patients with CR tend to do better than patients who achieved PR after salvage chemotherapy. To achieve this, patients with relapsed/refractory disease are currently treated with a variety of treatment regimens prior to them going for a transplant. Currently the most commonly used regimen is the chemotherapy regimen of ICE (ifosfamide-carboplatin-etoposide). Historically, when rituximab was added to this regimen (R-ICE), the number of patients responding increased. This increase was clinically significant. However, this could further be increased, since at present, this is estimated to be around 40% for patients who receive 2 cycles of therapy prior to them getting a transplant. </p><p> The goals of my master&rsquo;s program and thesis were, therefore, to (a) <b><u>develop rationale</u></b> for a clinical trial incorporating the novel drug lenalidomide into regimens treating patients with lymphoma (Chapter 1); (b) <b><u>review literature on paradigm changes</u></b> on how to treat patients with DLBCL in a molecular era (Chapter 2); (c) <b><u>secure funding and develop a clinical trial protocol</u></b> of the addition of lenalidomide to treat patients with DLBCL to the standard R-ICE regimen [R2ICE](Chapter 3); (d) to <b><u>report the early results of the safety from the completed Phase-1 study and ongoing phase-2 study for our R2ICE regimen</u></b>, and for its potential to become a <b><u>new regimen</u></b> for patients with relapsed/refractory lymphoma (Chapter 4). </p><p>