Cancer

Permanent URI for this collectionhttps://hdl.handle.net/20.500.12503/21704

Browse

Recent Submissions

Now showing 1 - 20 of 21
  • Item
    Activation of a putative membrane androgen receptor increases the efficacy of the chemotherapeutic agent, temozolomide, in a human glioblastoma cell line
    (2016-03-23) Badeaux-McGilvray, Adrienne; Singh, Meharvan PhD; Brock, Courtney
    Glioblastoma Multiforme (GBM) is a form of brain cancer with very poor prognosis such that the life expectancy of a person with this disease is about one year after diagnosis. Moreover, current treatment regimens are only able to extend the life span by mere months. Based on recent studies from our lab that identified a putative membrane androgen receptor (mAR), which when activated is capable of promoting cell death, we investigated whether exploitation of this receptor could increase the efficacy of current chemotherapeutic agents to combat this deadly and invariably lethal cancer. Using the human glioblastoma cell lines, A172 and T98G, our studies have shown that activation of the mAR (using testosterone or dihydrotestosterone conjugated to bovine serum albumin) not only sensitized the glioblastoma cells to temozolomide (TMZ), the current standard chemotherapeutic agent for GBM, but also suppressed the phosphorylation of Akt, a known survival-promoting factor. Further, in T98G cells that express high levels of O6-methylguanine DNA methyltransferase (MGMT), a DNA repair protein, activation of the mAR suppressed the expression of MGMT. Our data also suggest that these mechanisms may not be mutually exclusive such that inhibition of Akt phosphorylation in and of itself led to a reduction in MGMT expression. Collectively, our data support the targeting of a putative membrane androgen receptor as complementary treatment for glioblastoma.
  • Item
    Development and characterization of methylene blue-loaded nanoparticles for glioblastoma
    (2016-03-23) Ranjan, Amalendu; Vishwanatha, Jamboor; Castañeda-Gill, Jessica M.
    Glioblastoma (GBM) is the most common and aggressive primary brain tumor in adults over 45, resulting in an average survival of 15 months post-diagnosis and treatment. While recent research has provided essential information to aid diagnosis and treatment, GBM is known to cause relapse following traditional combinatorial regimens (surgery, radiation, and chemotherapy); this necessitates the development of more effective, less toxic therapies for diagnosed patients. Methylene blue (MB), a blue dye with noted medicinal applications, has received recent consideration as a potential neurotherapeutic due to its ability to infiltrate the blood-brain barrier (BBB), improve cellular processes within distinct brain cell compartments and types, and preferential accumulation in the brain. While MB displays these advantages, one drawback is an increased administration to produce desired therapeutic effects, leading to excessive brain deposition and potential neurotoxicity. A method commonly used to enhance drug delivery while reducing unwanted side effects is via encapsulation in submicron-sized nanoparticles (NPs) composed of the biodegradable/biocompatible co-polymer, poly(lactic-co-glycolic) acid (PLGA). Our lab, as well as others, have shown the application of PLGA NPs as potential cancer therapies, as well as their preferential accumulation in the brain, as a means to improve passive drug delivery. With this knowledge, our goal is to develop a MB-loaded PLGA NP capable of permeating the BBB in order to treat GBM, based on our hypothesis that encapsulation of MB into PLGA NPs will enhance accumulation in cancerous brain regions, resulting in reduced tumor size and prolonged survival. In this study, we formulated and characterized MB-loaded PLGA NPs via particle size, surface charge, drug loading, and encapsulation efficiency. Additionally, we analyzed their in vitro effects to establish biological, and potentially therapeutic, activity. Following MB loading and comparison to blank NPs, we obtained preparations comparable to those published sized at 162.4nm, a surface charge of -31.7, and drug loading and encapsulation efficiency values of 2.2% and 29.2%, respectively. With this data, MBNPs were further analyzed and determined to produce a peak release at 24hrs. Additionally, in in vitro cell death studies, MBNPs were found to induce cell death comparable to, if not better than, free MB, in GBM cell lines. Lastly, MBNP treatment enhanced cellular metabolism, an capability noted in free MB. We are currently completing additional studies on MBNPs’ effects in vitro, as well as establishing a protocol for a PK study. In conclusion, our formulation displays MB’s therapeutic potential, displayed by its enhanced effectiveness in in vitro studies compared to free MB.
  • Item
    Bone Microenvironment Targeted Nanoparticles for Metastatic Prostate Cancer Treatment
    (2016-03-23) Ranjan, Amalendu; Mukerjee, Anindita; Sarker, Marjana; Vishwanatha, Jamboor; Gdowski, Andrew
    Introduction The most common site of metastatic prostate cancer is the bone. These metastatic lesions are difficult to treat and often result in off target cytotoxicity from current chemotherapeutics. We hypothesize that targeted nanoparticles (NPs) designed to deliver chemotherapeutics to cancer lesions in the bone microenvironment could improve the side effect profile that results from non-discriminate action of cytotoxic agents. We have designed a novel targeted nanotherapeutic system to target the bone microenvironment in an effort to more efficiently delivery chemotherapeutics to the site of metastasis. The core of the NPs are composed of poly (D,L-lactic-co-glycolic acid) (PLGA) biodegradable polymer. The PLGA NPs have been loaded with the microtubule inhibitor, cabazitaxel. The surface of the NP has been conjugated with an amino-bisphosphonate, which has high affinity for the hydroxyapatite structure of the bone. Methods NPs were formulated using a modified water in oil in water double emulsion solvent evaporation technique. The physiochemical properties of the NPs were characterized. Ex vivo bone binding studies were performed. Cell viability studies were performed on C4-2B and PC3 cell lines. NPs were also tested on 3D tumor spheroids. Finally, NPs were tested for efficacy in an intraosseous tumor model of metastatic prostate cancer in athymic nude male mice. Results NPs were made with favorable physiochemical characteristics: mean hydrodynamic diameter of 236.8 nm ± S.D. 1.19, mean polydispersity of 0.121 ± SEM 0.003, encapsulation efficiency of 57%, and drug loading of 3.74. Cellular cytoxicity assay showed that C4-2B cells were more sensitive to the free cabazitaxel, the non-targeted NPs, and the targeted- NPs compared to PC-3 cells. We did not see an appreciable difference between the targeted-NPs and equivalent treatment of free cabazitaxel in 3D assays. In vivo analysis showed that both the non-targeted and targeted-NPs were more effective than free cabazitaxel at reducing tumor burden. Additionally, targeted-NPs improved bone morphology at tumor lesions. Conclusion In this project we have engineered a bone targeted NP formulation for metastatic prostate cancer. We have determined the chemical and physical characteristics of this system and tested the in vitro cytotoxicity. Finally, we have shown the efficacy of these targeted-NPs in an intraosseous model of bone metastatic prostate cancer.
  • Item
    Preventive Health Screenings in High Risk Populations: The Case of a HIV Positive Refugee Woman
    (2016-03-23) Raines-Milenkov, Amy; Lopez, Tania; Wurie, Neneh; Anderson, Ralph; Okaalet, Jenny
    Background: Women infected with HIV are 5.4 times more likely to be diagnosed with cervical cancer than women without HIV. It is recommended that women with HIV have a pap test at the time of HIV diagnosis and every three years after 3 consecutive Pap tests are normal. This case study will describe the chronology of key events from resettlement in the U.S. to a high grade abnormal PAP test result in an HIV positive refugee woman. Building Bridges Initiative (BBI) is a cancer education and screening program for refugee women at the UNT Health Science Center and CPRIT. The program trains and employs lay health educators from refugee communities to provide culturally and linguistically appropriate outreach, education and navigation services. Methods: A review of existing medical records, BBI case files, and medical case management (MCM) files explored multilevel factors associated with missed opportunities for pap screenings and education. Data was collected on cancer knowledge and PAP test awareness from self-reported baseline assessments. Results: Our case study is an HIV positive refugee from Africa who reported to have become HIV positive as a result of rape in 2007. The patient reported a previous PAP test in January 2014 prior to arrival in the U.S. (February, 2014). At BBI enrollment (February, 2015), she specifically expressed interest in a follow-up PAP exam. Upon completion of cancer education classes (3/24/2015), a Well woman/PAP test appointment was scheduled and completed on 6/26/2015 at UNTHSC-OBGYN clinic. Patient was informed of high grade abnormal results and referred (7/6/2015) to MCM for follow-up. She subsequently underwent a colposcopy and cold knife cone biopsy. Patient was receiving treatment for her HIV status since March of 2014. Although a PAP referral was noted in her records (12/8/2014) she did not receive a PAP exam in that system, but received one through the BBI 6 months later. Patients are required to make the PAP appointment themselves. Conclusion: Records indicate that the BBI participant kept all appointments for her medical management of HIV. Failure to receive a PAP exam in the same medical system may be due to the requirement that patients make their own appointments. Language and cultural barriers may have prevented her from doing so. Refugee women face trauma, such as rape and the possibility of HPV, HIV and other infectious diseases. Given their vulnerability and high risk, it may be prudent to provide PAP exams upon arrival.
  • Item
    Racial Disparities and Mortality Rates for Ovarian Cancer in Texas, 1999-2009
    (2016-03-23) Liu, Youcheng; Mehta, Pooja
    Background: Ovarian cancer is the leading cause of gynecological cancer-associated deaths, being the fifth most common of all cancer-associated deaths among women. Regional variation in mortality rates can be due to race and ethnicity, their distribution, varied reproductive patterns, tubal ligation, socioeconomic status, access to healthcare and diet. Other risk factors such as family history, early menarche, and late menopause, and nulliparity have also been identified. Hypothesis and Objective: The hypotheses of this study are: 1. Race is a predictor of mortality rates for ovarian cancer. 2. There is a significant difference in demographics, socioeconomic status and ovarian cancer mortality rates between 1999 and 2009 years in the counties of Texas. 3. Access to health care, age, lack of insurance, and reproductive pattern, are significant geographic variables associated with ovarian cancer mortality in Texas. The objective is to test these hypotheses. Materials and Methods: A cross-sectional study was conducted using the demographic and access to healthcare county level data obtained from the US Census Bureau 2000 and County Health Rankings and Roadmap website for the years 1999 and 2009 for women in Texas which were matched with the age-adjusted death rates (AADR) for the same periods obtained from the Texas Department of State Health Services and Texas State Data Center. Two sample t-tests were performed to evaluate the difference between the trends of 1999 and 2009. Correlation coefficient and multiple regression model analysis were performed to analyze the relationship between mortality rates and uninsured adults and between primary care provider rate (PPR) and socioeconomic status, respectively. Spatial analysis including mapping and hot spot analysis was also performed to find significant clusters. Results: Income level, education, and poverty levels improved significantly from 1999 to 2009 in white women but not in blacks. The average family size of whites (2.86) was significantly less than blacks (3.28). The PPR was negatively correlated to the mortality rates. Low median household income in past 12 months was significantly related to AADR (p-value= 0.01). Eastern Texas showed consistently high AADR from 1999 to 2009. Hot spot analysis revealed very few significant clusters in central and north Texas. Conclusion: Racial disparities and geographical factors can play an important role in predicting the disease outcome.
  • Item
    Outcomes Following Modern Treatment for Osteosarcoma: The Cook Children's Experience
    (2016-03-23) Bowman, Paul; Shah, Deep; Konty, Logan; Jones, Garrett; Akers, Lauren; Kiel, Alice; Allen, Joseph
    The purpose of this study is to evaluate patient and tumor characteristics, treatments, and outcomes of patients at Cook Children’s Medical Center (CCMC) who were diagnosed with primary osteosarcoma from January 1, 1992 to December 31, 2013. Prior to the use of systemic chemotherapy, the survival rate for osteosarcoma patients was poor at 15-20% for two year survival. Current five year survival is more than 60% in non-metastatic disease with the addition of systemic chemotherapy. However, patients with metastatic disease at diagnosis have a lower 5 year survival of about 30%. This study will evaluate the outcomes of patients with osteosarcoma at CCMC relative to the outcomes noted at other institutions and in previous literature. It will also evaluate various potential prognostic factors to determine if they significantly affect patient outcomes. Researchers conducted a comprehensive retrospective chart review with the patients abstracted from the Cook Children’s Cancer Registry. The population consists of patients diagnosed with and treated for primary osteosarcoma at CCMC from 1992-2013. The 5 year overall survival for the population is 66.9% ± 6.1. The 5 year Event Free Survival for the population is 62.2% ± 6.6. The 5 year survival for patients with metastases at diagnosis is 50% ± 13.9. The 5 year survival for patients without metastases at diagnosis is 72.3 % ± 6.8. The 5 year survival of patients diagnosed from 1995 to 2003 is 70.0% ± 8.9. The 5 year survival for patients diagnosed from 2004 to 2013 is 64.2% ± 8.8. Survival rates for patients diagnosed with osteosarcoma at CCMC are consistent with the literature. Survival rates have not changed in the past 20 years for osteosarcoma.
  • Item
    Radiotherapeutic Bandage for the Treatment of Skin Cancer
    (2016-03-23) Shi, Yi; Munaweera, Imalka; Zangla, Emily; Balkus, Kenneth Jr; Di Pasqua, Anthony PhD; Koneru, Bhuvaneswari
    Purpose: It is currently estimated that one in every five Americans will develop skin cancer. Squamous cell carcinoma (SCC) is the second most common type of skin cancer and occurs in cells just beneath the outermost layer of the epidermis. Radiation therapy is used in the clinic against inoperable tumor lesions and in patients that cannot undergo surgery, as well as to treat recurring lesions after a primary surgical approach (i.e., Mohs micrographic surgery). We have previously reported on the incorporation of Holmium-165 (165Ho) nanoparticles into electrospun nanofibrous mats (“bandages”) for potential use in the treatment of SCC. A 165Ho-containing polymer nanofibrous bandage was prepared via electrospinning using 165Ho-iron garnet nanoparticles (165HoIG) and polyacrylonitrile. These bandages can be manipulated for easy application to tumor lesions, and can be made on a large scale; they are made radioactive (to holmium-166; 166Ho) just prior to therapy using a process called neutron-activation. The goal of the present study is to test our radiotherapeutic bandage against SCC in an animal model, to determine clinical relevance. Methods: Polyacrylonitrile polymer bandages containing 165HoIG were prepared as previously reported. The radiotherapeutic bandages were then produced via neutron-activation in a thermal neutron flux of 1.8 × 1013 neutrons/cm2·s for 1.33 h using a 1 MW nuclear reactor. Female athymic nude mice were injected with human Colo-16 SCC cells subcutaneously and after eight days (average tumor volume: 35 ± 8.6 mm3) received no treatment, or were exposed to non-radioactive or radioactive (92.5 ± 18.5 MBq) bandages for approximately 1 h (n = 10 per group). After treatment, tumors were measured over fifteen days, tumor volume ratios (TVRs) compared and histopathology performed. Results: Fifteen days after treatment, the TVR of the radioactive bandage treatment group was 3.3 ± 4.5, while TVRs of the non-radioactive bandage treatment and no treatment control groups were 33.2 ± 14.7 and 26.9 ± 12.6, respectively. At the time of necropsy, there was mild focal epidermal hyperplasia surrounding a small area of epidermal ulceration in the radioactive bandage group. No other examined tissue (i.e., muscle, liver, kidney, lung, spleen and heart) showed significant lesions. Conclusions: Our radiotherapeutic bandage exhibits promising efficacy against SCC of the skin in a mouse model. It can be individually tailored for easy application on tumor lesions of all shapes and sizes, and could complement or possibly replace surgery in the clinic.
  • Item
    Racial Variation in Annexin A2 (AnxA2) Gene Expression and Poor Outcome in Triple-Negative Breast Cancer
    (2016-03-23) Vishwanatha, Jamboor; Gibbs, Lee
    Background: Triple-negative breast cancer (TNBC) is identified by the absence of three major receptors that drive most breast cancers and is a health disparity issue due to its disproportionate occurrence in African American (AA) women. Previous studies have identified elevated levels of AnxA2 in TNBC cell lines, but its association with racial variation and outcomes is unknown. Methods: AnxA2 gene expression was evaluated in TNBC and non-TNBC cases from The Cancer Genome Atlas (TCGA) RNAseqV2 database. Associations between clinical outcomes and AnxA2 gene expression were tested in a genome wide association study of combined publically available datasets. Results: AnxA2 gene expression was elevated in TNBC in comparison to other breast cancer subtypes. Furthermore, AnxA2 gene expression is elevated in AA women and is associated with the disproportionate occurrence of TNBC. High expression levels of AnxA2 is associated with reduced overall survival (hazard =2.66; 95% confidence interval {CI] = 1.14 - 6.25, P = 0.0192), reduced relapse free survival (hazard = 1.45; 95% confidence interval {CI] = 1.12 – 1.89, P = 0.0051), and reduced distant metastasis free survival (hazard = 1.7; 95% confidence interval {CI] = 1.00 – 2.91, P = 0.048). AnxA2 gene expression was not associated with poor outcome in other subtypes, indicating the specificity of AnxA2 contribution to the aggressive behavior of TNBC. Conclusion: AnxA2 overexpression is associated with racial variation and is a potential prognostic candidate for TNBC. AnxA2 has potential prognostic value, implicating a role for AnxA2 in the aggressive biology of TNBC in AA women.
  • Item
    Tolfenamic Acid and Curcumin Analogs Effectively Inhibits Pancreatic Cancer Cell Growth
    (2016-03-23) Basha, Riyaz; Sankpal, Umesh; Hurtado, Myrna
    Tolfenamic Acid and Curcumin Analogs Effectively Inhibits Pancreatic Cancer Cell Growth Background: Curcumin (Cur) is a natural phenol of the plant Curcuma longa that has been tested for anti-cancer activity in pre-clinical and clinical studies. The therapeutic efficacy of Cur is greatly impacted by its low bioavailability. Synthetic analogs of Cur are under testing to overcome this limitation. Cur analogs, EF31 and UBS109 have shown anti-cancer activity in colon and pancreatic cancers. Small molecule Tolfenamic acid (TA) is known to inhibit human cancer cells and tumor growth in mouse models for some human cancers. TA is known to target Specificity protein 1 (Sp1) transcription factor that mediates the expression of several genes associated with cancer including survivin, a key member of inhibitor of apoptosis proteins family. Purpose: Combination treatments have been tested to address the issues related to efficacy and drug resistance. In this project, the anti-cancer activity of EF31 and UBS109 was tested in combination with TA. Method: Human pancreatic cancer cells, L3.6pl and MIA PaCa-2 were grown in DMEM media with 5% fetal bovine serum. Cells were treated with DMSO (control) or EF31 or UBS109 or TA or EF31+TA or UBS109+TA and cell viability was measured at 48 hour post treatment using CellTiter-Glo (Promega) kit. The effect on apoptosis was assessed by determining the activity of caspase 3/7 (Caspase-Glo kit) and the expression of cleaved PARP (Western blot). The effect of individual and combination treatment(s) on the expression of Sp1 and survivin was evaluated by Western blot analysis. Results: TA in combination with EF31 or UBS109 resulted in an induction of cell growth inhibition which is accompanied by increased caspase 3/7 activity and upregulation of PARP cleavage. The combination treatment(s) also showed a modulation in the expression of Sp1 and survivin. Conclusions: TA combination with EF31 or UBS109 increases pancreatic cancer cell growth inhibition potentially by inducing the apoptotic pathways. When compared to TA, TA and Cur analog(s) is more effective in pancreatic cancer cells. These new combinations of anti-cancer molecules provide preliminary evidence for the therapeutic efficacy in treating pancreatic cancer.
  • Item
    Evaluating reconstituted high density lipoprotein nanoparticles as target specific Doxorubicin carriers using fluorescence spectroscopy.
    (2016-03-23) Requena, Sebastian; Sabnis, Nirupama; Chib, Rahul; Lacko, Andras G.; Borejdo, Julian; Gryczynski, Zygmunt; Gryczynski, Ignacy; Shah, Sunil Ajit
    Doxorubicin, also known as Adriamycin, is an anthracycline antibiotic which first gained clinical prominence in the early 1970’s as an effective antitumor agent. It is still used today to treat a spectrum of cancers like lymphoma, bladder, stomach, lung, breast, ovarian, and several others. Due to its production of free radicals to attack tumor cells, Doxorubicin interferes with mitochondrial phosphorylation and also induces cardiotoxicity. Thus, efficient and biocompatible delivery methods are needed for targeted drug delivery to overcome systemic toxicity. To maintain a high level of growth, tumor cells overexpress Scavenger receptors type B-1 (SR-B1). This cellular characteristic can be used to target and selectively deliver doxorubicin to tumor cells by packing it in reconstituted high density lipoprotein (rHDL) nanoparticles, which bind selectively to SR-B1 receptors. Nanoparticles as target-specific drug delivery agents are increasingly used in cancer therapy to enhance bioavailability and to reduce off target toxicity of anti-cancer agents. Several different formulations of rHDL nanoparticles to incorporate doxorubicin were synthesized. Doxorubicin’s intrinsic fluorescence was used to photophysically characterize the properties of loaded rHDL nanoparticles including absorption, emission, excitation, steady-state and time resolved anisotropy measurements, and quenching to study drug shielding by nanoparticles. Overall the fluorescence properties of the rHDL: doxorubicin complex may reveal valuable novel characteristics of this drug delivery vehicle that may be particularly applicable when used in systemic (intravenous) therapy.
  • Item
    In Vitro Study to Identify a Novel Combination Treatment for Ewing Sarcoma
    (2016-03-23) Sankpal, Umesh; Bowman, Paul; Ray, Anish; Conjeevaram Nagarajan, Bhavani Saranya; Basha, Riyaz; Gotlib, Daniel
    Introduction: Ewing sarcoma (ES) is an aggressive tumor that predominantly occurs in young adolescent populations. Chemotherapy used to treat ES is associated with long-term morbidity. The objective of this study was to identify an effective combination treatment option for treating ES. A fusion protein EWS-FLI1 is associated with [greater than] 85% of ES incidences and mithramycin (Mit) is known to target this fusion protein and is currently in clinical trials. We investigated the combination of Mit to induce the efficacy of Doxorubicin (Dox), a widely used chemotherapeutic agent for treating ES patients. Methods: ES cell lines CHLA9, CHLA10, and TC71 were cultured in laboratory and treated with increasing concentration of Mit or Dox. Cell viability was assessed using CellTiter-Glo luminescent assay at 48 hour post-treatment. Optimized doses for Mit and Dox were used for combination treatment. Western blot analysis was used to evaluate the apoptotic markers, caspase 3, cleaved PARP and Bcl2. Real-Time PCR was used to evaluate the expression of EWS-FL1 downstream targets. RNA was extracted using Trizol reagent (Invitrogen) and subjected to cDNA synthesis using Superscript III reverse transcriptase. PCR was carried out with cDNA using TaqMan primer-probes specific for ID2, LDB2, NROB1 and NCOR1. Results: The results of current study illustrated that Mit or Dox treatment resulted in a dose and time dependent decrease in ES cell proliferation. Mit down-regulated the EWS-FLI1 downstream targets (ID2, LDB2, NROB1 and RCOR1) in TC71 cells. Combination of Mit+Dox enhancesd the anti-proliferative effect in ES cell lines which was accompanied by modulation of apoptotic markers. Conclusion: This investigation provides evidence for the use of Mithramycin for enhancing the efficacy of chemotherapeutic agents. In vivo assays are underway to confirm in vitro results. These pre-clinical studies will have clinical implications for treating Ewing Sarcoma patients. The proposed combination to induce chemotherapy response is highly helpful since it can be tested to reducing chemotherapy dose(s) and addressing the issues related to side-effects.
  • Item
    A fluorescent biosensor for the detection and imaging of a cancer biomarker
    (2016-03-23) Mummert, Mark; Bora, Ilkay; Shah, Sunil; Gryczynski, Ignacy; Borejdo, Julian; Gryczynski, Zygmunt; Fudala, Rafal; Chib, Rahul
    Purpose: Cancer is among the leading cause of death worldwide with approximately 8.2 million deaths. Cancer mortality can be reduced if it can be detected and treated at early stages. Certain biomarkers like hyaluronidase are reported in the literature for the screening and detection of cancer. It is an endoglycosidase that degrades hyaluronic acid (HA) and this enzyme is overexpressed in various cancers. Therefore, it is of great interest to develop a simple, sensitive and fast technique with which one can estimate the activity/level of hyaluronidase. We have developed a fluorescent biosensor for the detection of hyaluronidase activity/level. This probe was developed by labeling hyaluronic acid with a long lifetime fluorophore. Monitoring the cleavage of hyaluronic acid by measuring the changes in the fluorescent properties of the biosensor will be a simple and precise tool reflecting hyaluronidase activity and can be used for the detection of cancer. Methods: This fluorescent biosensor was developed by heavy labeling of hyaluronic acid with an orange/red emitting azadioxatriangulenium (ADOTA) fluorophore. ADOTA in water emits at 560 nm with a long fluorescence lifetime of ~ 20 ns. The hyaluronidase activity was measured as a function of the change in the steady-state fluorescence intensity and fluorescence lifetime of the biosensor. Fluorescence lifetime imaging microscopy (FLIM) was also used to image hyaluronidase activity in cancer cells. Results: The heavily ADOTA labeled hyaluronic acid (HA-ADOTA) has a red shift in the peak emission wavelength (605 nm), a weak fluorescence signal and a short fluorescence lifetime due to efficient self-quenching. In the presence of enzyme hyaluronidase, the brightness and fluorescence lifetime of the sample increases with a shift in the peak emission to its original wavelength at 560 nm. The ratio of the fluorescence intensity of the HA - ADOTA probe at 560 nm and 605 nm can be used as the sensing signal for detecting hyaluronidase. Recovery in the fluorescence lifetime was used for fluorescence lifetime-based imaging of cancer. Conclusions: Our results show the ability of this the HA-ADOTA probe to detect activity/level of hyaluronidase in biological samples. Due to long fluorescence lifetime of the dye, it can be also be used to remove the background in cellular images. In future, this novel technology can be used to design a small device which can be used in primary care settings for the screening of cancer.
  • Item
    Hematopoietic Stem Cell/Bone Marrow Transplant in Pediatric Acute Lymphoblastic Leukemia: A Retrospective Analysis of Survival Rate and Treatment-related Mortality
    (2016-03-23) Kiel, Alice; Bowman, Paul; Hernandez, Joshua
    Purpose: Acute Lymphoblastic Leukemia (ALL) makes up nearly 75% of childhood leukemias, making it the most common pediatric malignancy. In cases where refractory disease occurs, allogeneic hematopoietic stem cell transplantation is the therapy recommended. Due to limited information on long-term clinical outcome of transplant recipients in each remission status, the results of this study may help identify a possible timely approach for myeloablative transplant for high risk ALL patients. Additionally, this report describes the difference in survival rates for T cell and B cell ALL populations. Methods: Researchers conducted a comprehensive retrospective chart review of patients identified from the Cook Children's Medical Center (CCMC) Cancer Registry. The population consists of patients diagnosed with ALL at CCMC from January 1, 1986 to December 31, 2012, who underwent a Stem Cell/Bone Marrow transplant. Statistical analysis of the data was performed to determine the survival rates and what factors may have statistically been associated with survival or with death due to the disease or treatment complications, as well as specifically comparing B and T cell ALL survival rates. Results: Overall, CCMC ALL patients who received a bone marrow transplant were found to have a five-year survival rate of 61.3% ± 3.6%, and a ten-year survival was 48.4 % ± 3.9 %. Evaluating the B cell ALL patients separately, the five and ten-year survival rates were 66.7% ± 3.8% and 52.6% ± 4.2%, respectively. The T cell patients when evaluated separately revealed five and ten-year survival rates of 29.5% ± 9.0% and 23.6% ± 8.9%, respectively. Conclusion: The survival rates for CCMC ALL patients who received bone marrow transplants were determined in this study. The data, when looked at separately, showed that there is a statistically significant difference between B cell and T cell survival rates as was hypothesized. T cell ALL survival rates were statistically significantly less than those of B cell ALL cases undergoing transplant. This information is important in helping us to better address the current treatment recommendations for ALL patients and objectively evaluate the effectiveness of those recommendations. The data collected has also provided the opportunity to further evaluate other related factors, such as Philadelphia chromosome and changes in mortality rates over time, and the effect they may have on long-term survival of ALL patients.
  • Item
    Focal Nodular Hyperplasia (FNH) of the Liver Following Treatment of Childhood Cancer
    (2016-03-23) Bowman, Paul; Cope, Sandy; Hanson, Gregory
    Purpose: The purpose of this case study is to describe three different patients with complicated malignancies in their past medical history who received the diagnosis of FNH. The report will highlight the clinical features of each case, describe the diagnostic modalities utilized and increase awareness of FNH as a long term sequela of chemotherapy and radiotherapy used to treat childhood cancers. Methods: Medical records of three patients treated at Cook Children’s Medical Center who were identified to have FNH were collected. Medical history, medications, clinician notes, imaging, surgical reports, and pathology reports were all reviewed systematically. A literature review was conducted using PubMed, Scopus and UpToDate. Results: Two of the three patients had been successfully treated for metastatic and recurrent Wilms’ tumors; the third had been successfully treated for embyronal rhabdomyosarcoma of the pelvis. All three patients required chemoradiation leading to extensive follow up for the long term side effects of their therapy. All three cases of FNH were found incidentally on imaging, which was either routine or based on symptoms unrelated to the liver. The lesions were first identified an average of 9 years after completion of cancer therapy. Two of the patients underwent surgical biopsy while the third was diagnosed based only on imaging. A watchful waiting approach was applied to all three children and they are all doing well without any hepatic complications, although the lesions often continue to increase in size slowly. Conclusions: The pathogenesis of FNH is thought to be due to a vascular insult leading to ischemia and subsequent remodeling of the hepatic tissue with reperfusion. It is commonly found in adult women on estrogen therapy. The exact mechanism of its development in children is unknown; however, there appears to be a correlation with a past medical history of childhood malignancy and chemoradiation therapy. It is possible to make the diagnosis based on imaging alone to avoid more invasive diagnostic procedures, if the lesions are characteristic and if they are not suspicious for metastatic or recurrent disease. A watchful waiting approach is recommended in all cases of focal nodular hyperplasia of the liver.
  • Item
    Bone Microenvironment Targeted Nanoparticles for Metastatic Prostate Cancer Treatment
    (2016-03-23) Ranjan, Amalendu; Mukerjee, Anindita; Sarker, Marjana; Vishwanatha, Jamboor; Gdowski, Andrew
    Introduction The most common site of metastatic prostate cancer is the bone. These metastatic lesions are difficult to treat and often result in off target cytotoxicity from current chemotherapeutics. We hypothesize that targeted nanoparticles (NPs) designed to deliver chemotherapeutics to cancer lesions in the bone microenvironment could improve the side effect profile that results from non-discriminate action of cytotoxic agents. We have designed a novel targeted nanotherapeutic system to target the bone microenvironment in an effort to more efficiently delivery chemotherapeutics to the site of metastasis. The core of the NPs are composed of poly (D,L-lactic-co-glycolic acid) (PLGA) biodegradable polymer. The PLGA NPs have been loaded with the microtubule inhibitor, cabazitaxel. The surface of the NP has been conjugated with an amino-bisphosphonate, which has high affinity for the hydroxyapatite structure of the bone. Methods NPs were formulated using a modified water in oil in water double emulsion solvent evaporation technique. The physiochemical properties of the NPs were characterized.Ex vivo bone binding studies were performed. Cell viability studies were performed on C4-2B and PC3 cell lines. NPs were also tested on 3D tumor spheroids. Finally, NPs were tested for efficacy in an intraosseous tumor model of metastatic prostate cancer in athymic nude male mice. Results NPs were made with favorable physiochemical characteristics: mean hydrodynamic diameter of 236.8 nm ± S.D. 1.19, mean polydispersity of 0.121 ± SEM 0.003, encapsulation efficiency of 57%, and drug loading of 3.74. Cellular cytoxicity assay showed that C4-2B cells were more sensitive to the free cabazitaxel, the non-targeted NPs, and the targeted- NPs compared to PC-3 cells. We did not see an appreciable difference between the targeted-NPs and equivalent treatment of free cabazitaxel in 3D assays. In vivo analysis showed that both the non-targeted and targeted-NPs were more effective than free cabazitaxel at reducing tumor burden. Additionally, targeted-NPs improved bone morphology at tumor lesions. Conclusion In this project we have engineered a bone targeted NP formulation for metastatic prostate cancer. We have determined the chemical and physical characteristics of this system and tested the in vitro cytotoxicity. Finally, we have shown the efficacy of these targeted-NPs in an intraosseous model of bone metastatic prostate cancer.
  • Item
    Isolation and Characterization of a Group of TS-FK228 Analogues
    (2016-03-23) Cheng, Yi-Qiang; Liu, Xiangyang
    FK228 (Istodax/Romidepsin) is an FDA approved anticancer drug for the treatment of human cutaneous T-cell lymphoma and peripheral T-cell lymphoma via the inhibition of class I histone deacetylases (HDACs). Fermentation of Chromobacterium violaceum No. 968 is still the main preparation method for the large-scale production of FK228 for its research and clinical applications. Interestingly, we previously discovered the production of authentic FK228 by Burkholderia thailandensis MSMB43. During a pilot scale production of FK228 through the fermentation of B. thailandensis MSMB43, we unexpectedly isolated and purified one labile compound (thiosulfinate-FK228, TS-FK228, existed as isomers) which is structurally similar to FK228 but distinctive from their cytotoxic effects against tumor cells. Through LC-MS analysis and subsequent chemical synthesis of five TS-compounds from the prepared FK228 and FK228 analogues (thailandepsins, TDPs), we were able to prove that the oxidation of the disulfide bond in FK228 is derived from silica gel during the course of silica gel chromatography rather than through de novo biosynthesis by perspective bacterial strain. We showed that one of the TS-FK228 isomers has similar HDAC inhibitory activities with FK228, but has higher antiproliferative activities than FK228 against representative human cancer cell lines.
  • Item
    Nitric oxide- and cisplatin-releasing amine-modified mesoporous silica nanoparticles for the treatment of non-small cell lung cancer
    (2016-03-23) Munaweera, Imalka; Koneru, Bhuvaneswari; Patel, Amit; Dang, Mai; Balkus, Kenneth Jr; Di Pasqua, Anthony PhD; Shi, Yi
    Purpose: Lung cancer is the leading cause of cancer related death in the United States; non-small cell lung cancer (NSCLC) is the most common type and it is a challenge to treat in the clinic. Studies have shown that cisplatin-loaded nanoparticles are efficacious against NSCLC and can reduce the nephrotoxicity associated with cisplatin. Previously, nitric oxide (NO) was shown to enhance the efficacy of chemo- and radiotherapies in vivo; thus, the aim of this study was to develop a NO- and cisplatin-releasing wrinkle-structured amine-modified mesoporous silica (AMS) nanoparticle for improved NSCLC therapy. Methods: The AMS and NO- and cisplatin-loaded AMS materials were prepared and characterized. Platinum (Pt) drug loading was analyzed using inductively coupled plasma-mass spectrometry (ICP-MS), and NO release and in vitro release of cisplatin from AMS materials were investigated. Cytotoxicity of functionalized AMS nanoparticles were tested against human NSCLC cell lines, H596 and A549, and compared with that against normal lung cell lines, WI-38 and BEAS-2B. Pt cellular uptake studies were then performed using these four cell lines. The results were then compared to those obtained for cisplatin. Results: For both NSCLC cell lines, the toxicity of NO- and cisplatin-loaded silica nanoparticles (NO-Si-DETA-cisplatin-AMS) was significantly higher than that of silica nanoparticles loaded with only cisplatin (Si-DETA-cisplatin-AMS). In contrast, the toxicity of NO-Si-DETA-cisplatin-AMS toward normal lung cell lines was not significantly different from that of Si-DETA-cisplatin-AMS (normal lung fibroblast cells WI-38) or was lower than that of Si-DETA-cisplatin-AMS (normal lung epithelial cells BEAS-2B). The calculated therapeutic index of NO-Si-DETA-cisplatin-AMS was higher than that of Si-DETA-cisplatin-AMS and free cisplatin, based on both WI-38 and BEAS-2B data. When treated with Si-DETA-cisplatin-AMS, Pt cellular uptake was highest in BEAS-2B, then H596, WI-38 and A549, which corresponds to the toxicity data, and Pt uptake was higher in NSCLC cells when treated with NO-Si-DETA-cisplatin-AMS, compared to Si-DETA-cisplatin-AMS; however, differences in Pt uptake were not statistically significant. Conclusions: The NO-induced sensitization of tumor cell death demonstrates that NO is a promising enhancer of platinum-based lung cancer therapy, and our new therapy may be useful to enhance efficacy of Pt therapy in the clinic.
  • Item
    Colorectal cancer mortality is decreasing but not the average person-years of life loss: Findings from a population-based study
    (2016-03-23) Nandy, Rajesh; Suzuki, Sumihiro PhD; Mamun, Md Abdullah
    Purpose: Colorectal cancer death rate decreased by about 2 percent per year in the 1990s and this decline accelerated to about 3 percent per year during 2001 to 2010 in the US. The objective of this study was to investigate whether the average person-years of life loss (APYLL) has improved in a similar fashion over time. We also investigated the effects of cancer stage at diagnosis on person-years of life loss (PYLL) since it is one of the major factors that influences the survival of cancer patients. Methods: A total of 509,738 patients diagnosed with only colorectal cancer in their lifetime in 1988-2012 were identified from the Surveillance, Epidemiology, and End Results (SEER) registries. We used life expectancy from the US population life table adjusted by year of death, sex, and race to estimate person-years of life loss. PYLL were calculated for 111,704 patients, who died during the follow up period due to colorectal cancer, by subtracting age at death from expected years of life at that age. To estimate the effect of early diagnosis on PYLL by controlling the effects of sex, race, and marital status, we have fitted a multiple linear regression. Results: Average PYLL was 16.3 years and showed an increasing trend from 14.6 years in 1988 to 17.4 years in 2012. Effects of stage at diagnosis on PYLL were significantly different between stages, and it varied from 10.8 years at stage 0 to 18.2 years at stage IV. Estimated APYLL for White, Black, and Other races were 15.5, 17.4, and 17.8 years respectively. After adjusting sex, race, and marital status we had found that asymptomatic diagnosis of colorectal cancer (stage 0) could save, on an average, 6.8 years of life loss in comparison to diagnosis at stage IV. Diagnosis at stage I, II, and III could save, on an average, 5.5, 4.3, and 2.3 years of life loss, respectively, in comparison to stage IV. Conclusion: Average person-years of life loss has been increasing steadily over the last two decades for colorectal cancer patients in the US. This increasing trend might be attributed to diagnosis at later stage and transition of the disease at younger age. It is evident from our study that early diagnosis might be one of the most effective ways to save person-years of life loss due to colorectal cancer.
  • Item
    Factors affecting awareness of Hepatitis B status among Bhutanese, Karen, Somali, and Central African Refugee populations in Tarrant County: Building Bridges Initiative (BBI)
    (2016-03-23) Felini, Martha; Raines-Milenkov, Amy; Baker, Eva; Okaalet, Jenny
    Background: Hepatitis B Virus is reported to be the leading cause of liver cancer in the United States, and 90% are foreign born. The CDC recommends HBV screening for newly arrived refugees who have lived in countries with a high prevalence of chronic HBV infection. Nearly 14,000 refugees resettled in Texas in 2014. Currently, limited data is available on refugee awareness of HBV. This study aims to investigate what factors influence their awareness of HBV status. Methods: Four community health workers conducted outreach and baseline assessments in their respective communities (i.e. Bhutanese, Burmese, Somali and Central African). Means, t-tests, and chi-square tests evaluated the influence of time in the US, education, and group on awareness of self-reported Hepatitis B status at baseline. Results: Approximately 350 women have participated in BBI. 60% had heard of hepatitis B. Only 26% were aware of their Hepatitis B status. The bivariate analysis showed nearly half of the central African women were aware for their HBV status (47.3%) in comparison with Somali (30.8%), Burmese (18.7%), Bhutan (3.3%). On average, participants lived in the US for approximately 5 years. Education and region were not significantly associated with awareness of HBV status. Conclusion: Majority (74%) of refugees are unaware of their Hepatitis B status. Time in the U.S., nor formal education influence HBV status awareness. However, awareness between regions shows that the differences within ethnic groups (traditions, health care beliefs) need to be considered. Refugee populations remain in need of culturally and linguistically appropriate cancer prevention programs.
  • Item
    Tolfenamic Acid Sensitizes Ewing Sarcoma Family Tumor Cells to Chemotherapy
    (2016-03-23) Sankpal, Umesh; Mike-Mayer, Austin; Tabor-Simecka, Leslie; Bowman, W. Paul; Ray, Anish; Basha, Riyaz; Shelake, Sagar
    Ewing sarcoma family tumor (EFT) is the second most prevalent bone and soft tissue tumor observed in children and young adolescents. Patients with metastatic disease have poor outcome with 5-year overall survival rate less than 30%. Current chemotherapeutic options are causing limited progress in the management of EFT. Our aim was to identify novel targets and less toxic agents to improve the efficacy of standard care offered to EFT patients. Specificity protein 1 (Sp1) transcription factor is known to be upregulated in various cancers and is frequently linked to poor prognosis. In this study, Tolfenamic acid, an inhibitor of Sp1 was tested to sensitize EFT cells to commonly used chemotherapeutic agent Vincristine (Vin). The effect of TA or Vin or TA+Vin on cell viability was evaluated by CellTiter-Glo assay and caspase 3/7 activity was measured by Caspase3/7 Glo assay. Western blot analysis was performed to determine the expression of Sp1, survivin, and cleaved-PARP. Apoptotic cell population was measured by Annexin V staining. Results showed a dose and time dependent decrease in cell viability with both agents while the combination of TA+Vin caused a significantly higher response as compared to individual treatments. This inhibition of cell viability was accompanied by the inhibition of Sp1 and survivin expression and an increase in the apoptotic markers i.e. Annexin-V positive cells, caspase 3/7 activity and the levels of c-PARP. Our results suggest that TA+Vin combination treatment provides an effective therapeutic strategy for the treatment of EFT.