Chemotherapeutic Administration Research Articles

Investigating the Potential of Extracellular Vesicles as Delivery Systems for Chemotherapeutics

Background/Objectives: Standard chemotherapy is generally considered the best approach to treat many solid cancers, even accounting for severe side effects. Therefore, the development of a drug delivery system for chemotherapeutic administration could significantly improve standard chemotherapy by maintaining the cytotoxic effects of the drugs while decreasing the inherent side effects of the treatment. The aim of our study is the optimization of a loading strategy that conjugates the use of extracellular vesicles (EVs) as drug delivery carriers, by preserving their integrity, with the loading efficiency and activity maintenance of chemotherapeutics. Methods: We compared the EV loading of the chemotherapeutics epirubicin, mitomycin, methotrexate and mitoxantrone by co-incubation. Once loaded, the activity of drug-carrying EVs was tested on cancer cells and compared to that of free chemotherapeutics. Results: We defined a linear correlation between chemotherapeutics’ concentration and their absorbance at the drug-specific wavelength, which allowed the definition of a highly sensitive absorbance-based spectrophotometric quantification system, enabling the assessment of drug loading efficiency. Co-incubation of EVs and chemotherapeutics was sufficient to obtain quantifiable drug loading, and the efficacy of EV loading was drug-dependent. Epirubicin-loaded vesicles showed increased toxicity to bladder cancer cells with respect to the free chemotherapeutic. The cytotoxicity was maintained even upon 6-month storage at −80 °C of loaded EVs. Conclusion: We established an absorbance-based spectrophotometric quantification system that enables a straightforward measure of drug loading efficiency into EVs, and we demonstrated that chemotherapeutic-carrying EVs can be obtained by co-incubation, preserving and increasing drug cytotoxicity.

Dual drug co‐encapsulation of bevacizumab and pemetrexed clocked polymeric nanoparticles improves antiproliferative activity and apoptosis induction in liver cancer cells

AbstractNanoparticle (NP) enabled approaches have been employed for chemotherapeutic administration due to their capacity to regulate drug release and reduce side effects. Additionally, these methods can use several drugs concurrently and impede the proliferation of cancer cells that have developed resistance. Bevacizumab (BVZ) and pemetrexed (PEM) have demonstrated encouraging outcomes in the treatment and management of cancer. This work investigates the combined antiproliferative efficacy of BVZ and PEM co‐loaded PLGA‐PEG NPs (BVZ/PEM@PLGA‐PEG NPs) against HepG2 liver cancerous cells. The BVZ/PEM@PLGA‐PEG exhibited a sphere‐shaped and consistent nanosized distribution. In addition, we evaluated the potential mechanisms for inhibiting cell growth and inducing apoptosis using DAPI staining and cell cycle study. The beneficial combined antiproliferative activity and the apoptosis pathway were detected in the HepG2 cells exposed to BVZ/PEM@PLGA‐PEG NPs. Our study determined that the combinational drug treatment of BVZ/PEM@PLGA‐PEG NPs has a significant effect on promoting the effectiveness of liver cancer treatment.

Skin Cancer Treatment with Subcutaneous Delivery of Doxorubicin-Loaded Gelatin Nanoparticles and NIR Activation.

Subcutaneous (SC) administration of chemotherapeutics combined with near-infrared (NIR) light activation can effectively target skin tumors by triggering localized drug release and enhancing cytotoxic effects. In this study, we developed NIR-responsive indocyanine green (ICG) and the chemotherapeutic agent doxorubicin (Dox) loaded into gelatin nanoparticles (NPs) for SC delivery in a skin tumor-bearing mouse model. Histological examination (hematoxylin and eosin staining) confirmed the successful delivery and swelling behavior of the Dox/ICG-loaded gelatin NPs at the SC site. In vitro and in vivo experiments demonstrated that NIR activation of the Dox/ICG-loaded gelatin NPs generated significant photothermal heat (48 and 46 °C), leading to targeted drug release and a substantial reduction in skin tumor size (from 15 to 3 mm3). Our findings suggest that this dual-modality approach of SC chemotherapeutic administration and NIR-triggered photothermal therapy can concentrate cytotoxic drugs at the tumor site, offering a promising strategy for improving skin cancer treatment.

Combined therapy with methotrexate nanoconjugate and dendritic cells with downregulated IL-10R expression modulates the tumor microenvironment and enhances the systemic anti-tumor immune response in MC38 murine colon carcinoma.

Understanding the negative impact of the tumor microenvironment on the creation of an effective immune response has contributed to the development of new therapeutic anti-cancer strategies. One such solution is combined therapy consisting of chemotherapeutic administration followed by dendritic cell (DC)-based vaccines. The use of cytostatic leads to the elimination of cancer cells, but can also modulate the tumor milieu. Moreover, great efforts are being made to increase the therapeutic outcome of immunotherapy, e.g. by enhancing the ability of DCs to generate an efficient immune response, even in the presence of immunosuppressive cytokines such as IL-10. The study aimed to determine the effectiveness of combined therapy with chemotherapeutic with immunomodulatory potential - HES-MTX nanoconjugate (composed of methotrexate (MTX) and hydroxyethyl starch (HES)) and DCs with downregulated expression of IL-10 receptor stimulated with tumor antigens (DC/shIL-10R/TAg) applied in MC38 murine colon carcinoma model. With the use of lentiviral vectors the DCs with decreased expression of IL-10R were obtained and characterized. During in vivo studies MC38-tumor bearing mice received MTX or HES-MTX nanoconjugate as a sole treatment or combined with DC-based immunotherapy containing unmodified DCs or DCs transduced with shRNA against IL-10R (or control shRNA sequence). Tumor volume was monitored during the experiment. One week after the last injection of DC-based vaccines, tumor nodules and spleens were dissected for ex vivo analysis. The changes in the local and systemic anti-tumor immune response were estimated with the use of flow cytometry and ELISA methods. In vitro studies showed that the downregulation of IL-10R expression in DCs enhances their ability to activate the specific anti-tumor immune response. The use of HES-MTX nanoconjugate and DC/shIL-10R/TAg in the therapy of MC38-tumor bearing mice resulted in the greatest tumor growth inhibition. At the local anti-tumor immune response level a decrease in the infiltration of cells with suppressor activity and an increase in the influx of effector cells into MC38 tumor tissue was observed. These changes were crucial to enhance the effective specific immune response at the systemic level, which was revealed in the greatest cytotoxic activity of spleen cells against MC38 cells.

Intra-arterial regional chemotherapy for pancreatic cancer (review)

Pancreatic cancer, despite its low prevalence, is the fourth most common cause of cancer death, with a 5-year survival rate of no more than 10 %. Experts predict that by 2030, pancreatic cancer will become the second most common cause of death from malignant neoplasms, surpassing colorectal and breast cancers. Thus, pancreatic cancer is characterized by very unfavorable prognosis, primarily due to the chemoresistant nature of the tumor.Only a few treatment options for pancreatic cancer are currently available, with low response rates, short progression-free survival and short overall survival, and severe toxicity. In order to improve clinical outcomes, a number of studies on the use of regional chemotherapy as a treatment option for pancreatic cancer have demonstrated dose-dependent tumor sensitivity. Chemotherapy by intra-arterial perfusion of the pancreas made it possible to increase local concentrations of chemotherapeutic agents with minimal effect on healthy tissues and a lower incidence of side effects compared to systemic chemotherapy. This therapeutic approach has demonstrated a good therapeutic effect in the treatment of malignant neoplasms of other localizations. This review summarizes clinical approaches to chemotherapeutic administration by intra-pancreatic intra-arterial perfusion in terms of methods, pharmacokinetics, and clinical outcomes.

De Ritis Ratio and related Parameters among Breast Cancer Patients in a Southern Nigeria Population

De Ritis described the ratio between the serum levels of liver enzymes; aspartate transaminase (AST) and alanine transaminase (ALT). The central role of the liver in metabolism makes it a necessity that it functions optimally during chemotherapeutic administration, hence the consideration of associated parameters in the management of breast cancer. This study was conducted in University of Calabar Teaching Hospital, Calabar. It included 40 cases of pathologically diagnosed breast cancer female patients who were accessing post-operative chemotherapy. Another 40 apparently healthy females drawn from the general population served as controls. Ethical approval and written informed consent were duly sought and obtained. The present study measured the levels of total and conjugated bilirubin as well as the enzyme activities for a spartate aminotransferase, alanine aminotransferase and alkaline phosphatase. Additionally, Unconjugated bilirubin derived from total and conjugated bilirubin as well as De Ritis ratio computed from aspartate aminotransferase and alanine aminotransferase were calculated. Standard colorimetric methods were used to determine the measured parameters, while derived parameters were calculated. The data obtained were analyzed in Statistical Package for Social Sciences (SPSS) using students t-test at 95% confidence level with p-value of ≤ 0.05. Apart from alanine aminotransferase that of the chemotherapy courses showed some observable effect on both derived parameters; Unconjugated Bilirubin and De Ritis ratio. The later showed a stable pattern of increment after commen cement of post-operative chemotherapy. This study indicates that routine assessment of hepatic function and De Ritis ratio should be determined as part of breast cancer management.

Reducing paclitaxel hypersensitivity reactions in gynecologic oncology patients (521)

Reducing paclitaxel hypersensitivity reactions in gynecologic oncology patients (521)

Alleviation of chemotherapy-induced acute lung injury via NLRP3/ ASC/ Caspase-1 signaling pathway.

Acute lung injury has been reported following various chemotherapeutic agents administration. Several pathways for lung injury have been speculated however, the exact mechanism of the lung injury induced by methotrexate (MTX) is yet to be defined. The potential protective effect of Ginkgo biloba extract (GB), a Chinese herbal medicine, against MTX-induced lung injury is still not reported. Therefore, this study was performed to examine the possible implication of NLRP3 inflammasome and miRNA-21 in the pathogenesis of the MTX-induced lung injury as well as the protective role of GB in ameliorating the induced lung injury. Rats received GB (100mg/kg/day, orally) for 10days and MTX (20mg/kg single dose, intraperitoneally) on the fifth day. MTX-induced lung injury was manifested by lung histopathology. MTX exhibited a marked increase in lung malondialdehyde beside a notable decrease in lung reduced glutathione. Moreover, MTX injection activated the lung NLRP3 inflammasome by significant upregulation of the NLRP3, ASC, caspase-1 lung mRNA expressions and protein levels in addition to lung NF-kBp65 protein expression, and miRNA-21 expression when compared with the normal control group. However, GB administration mitigated lung injury and inhibited the NLRP3 activation. This study is the first report to reveal the involvement of NLRP3 inflammasome in the pathogenesis of MTX-induced lung injury and also to show that the administration of GB alleviates the lung injury induced by MTX via suppressing the oxidative stress, restoring the antioxidant activity, blocking the NLRP3/ASC/Caspase-1 signaling and downregulating the NF-kBp65 protein expression ae well as miRNA-21 expression in lung tissue.

Computational modeling of schedule-specific chemotherapy outcomes in mouse tumor model

Despite the rapid development of new innovative strategies in cancer treatment like immunotherapy, chemotherapy still remains a common choice in many cases. Standard protocols of chemotherapeutic administration rely on a maximal tolerated dose paradigm, however there is growing evidence that this approach is not always optimal. Alternative scheduling, like metronomic - low dose continuous drug administration - were recently proved their efficacy. The space of available variants of drug administration protocols is prohibitively large to be explored empirically, and there is an urgent need of predictive mathematical models for rational chemotherapeutic scheduling design. In this work we tested the ability of the minimal pharmacokinetic-pharmacodynamics model to describe schedule-specific tumor volume time evolution in different mouse tumor models.

Cancer Chemotherapy: A Review Update of the Mechanisms of Actions, Prospects, and Associated Problems

Cancer is one of the leading causes of death in developed countries, responsible for about 25% of all deaths. On a yearly basis, 0.5% of the population is diagnosed with cancer. Treatment options include surgery, radiotherapy, and systemic therapies such as chemotherapy, endocrine therapy, and targeted agents. Conventional chemotherapeutical agents act by creating toxic effects on all dividing cells. The toxic by-products of some chemotherapeutic drugs are deposited mainly in the liver producing hepatotoxicity, leading to many side-effects like loss of appetite, nausea, vomiting, diarrhea, abdominal cramps and constipation, anemia, fatigue, hyperthermia, loss of hair, etc. In this review update, the benefits and problems of chemotherapy have been highlighted and the mechanisms involved in cancer formation, groups of chemotherapeutic agents, as well as method of chemotherapeutic administration, have been discussed

The combined role of NT-proBNP and LV-GLS in the detection of early subtle chemotherapy-induced cardiotoxicity in breast cancer female patients

BackgroundChemotherapeutic agents have many side effects; among them is cardiotoxicity. Ejection fraction fails to detect the subtle alterations of left ventricular (LV) function; that is why there is a need for a more sensitive tool. The aim is to detect subclinical LV systolic dysfunction after chemotherapeutic treatment, using NT-BNP plasma level as well as speckle tracking echo-global longitudinal strain (STE-GLS). Seventy-four asymptomatic, non-metastasizing breast cancer female patients without risk factors were included. They were assessed before and 6 weeks after taking their first chemotherapeutic session. Assessment included clinical characteristics, conventional two-dimensional (2D) and three-dimensional (3D) echocardiography, and 2D STE-GLS. Blood samples for NT-BNP plasma level were collected on both visits and were later analyzed using a Sandwich ELISA technique.ResultsThe median NT-proBNP almost doubled after 6 weeks of chemotherapy (73.50 vs 34.4 pg/L, p value <0.001). Only two patients showed significant reduction of LVEF >10% to less <55%. One patient died before her scheduled follow-up visit, and the cause of death is unknown. Fifty patients showed elevated follow-up levels of the NT-BNP. As compared to the baseline visit, 12 patients had a high relative reduction of the LV-GLS (>15%) and all of them had a relatively higher NT-proBNP. A 2.2 relative elevation of the NT-proBNP was able to define a relative reduction of LV-GLS >15% by a 100% sensitivity and 81.8% specificity.ConclusionThe relative reduction of LV-GLS and the relative elevation of NT-proBNP were successful in defining subclinical, subtle chemotherapy-induced cardiotoxicity after 6 weeks of the first chemotherapeutic agent administration.

Abstract PO065: Single cell transcriptomics of triple negative breast cancer allografts following chemotherapy treatment reveals increased T cell abundance in doxorubicin-sensitive tumors

Abstract The contribution of stromal cells on drug response in primary tumors remains unclear. To determine how individual cells within the stroma of triple negative breast cancer (TNBC) allografts respond to chemotherapy, we used single cell sequencing to profile cells present in murine tumors with or without exposure to doxorubicin. Doxorubicin is a chemotherapeutic agent commonly used to treat TNBC by inhibiting cancer cell proliferation through intercalation of DNA and preventing topoisomerase II activity. In this study, murine TNBC 4T1 cell line was utilized to generate allograft tumors in immunocompetent BALB/c mice. Syngeneic 4T1 tumors exhibit a range of responsiveness to doxorubicin treatment. Tumor growth rates were monitored throughout the chemotherapeutic regiment then stratified into sensitive or resistant response. Cellular composition and behavior were then analyzed via single cell RNA sequencing (scRNA-seq) and flow cytometry 8 days post-doxorubicin chemotherapeutic administration mimicking clinical treatment. ScRNA-seq revealed decreases in tumor infiltrating lymphocytes after doxorubicin exposure. Furthermore, an increase in T cell abundance was discovered in tumors sensitive to doxorubicin treatment. This finding was further supported by flow cytometric analysis showing a tumor specific increase in all, CD4+, and CD8+ T cell populations relative to resistant tumors. Additionally, T-cell differentiation, exhaustion, and activation states were further examined from scRNA-seq data providing insights into functional properties of the tumor residing T cell populations undergoing chemotherapeutic treatment. Future work will focus on analyzing prognostic value of specific T-cell populations in disease regression following doxorubicin treatment. This study received funding from LLNL LDRD grant 19-SI-003. This work was conducted under the auspices of the USDOE by LLNL (DE-AC52-07NA27344). Citation Format: Nicholas R. Hum, Aimy Sebastian, Sean F. Gilmore, David M. Gravano, Naiomy D. Rios-Arce, Kelly A. Martin, Elizabeth K. Wheeler, Matthew A. Coleman, Gabriela G. Loots. Single cell transcriptomics of triple negative breast cancer allografts following chemotherapy treatment reveals increased T cell abundance in doxorubicin-sensitive tumors [abstract]. In: Abstracts: AACR Virtual Special Conference: Tumor Immunology and Immunotherapy; 2020 Oct 19-20. Philadelphia (PA): AACR; Cancer Immunol Res 2021;9(2 Suppl):Abstract nr PO065.

Differential effect on different immune subsets of neoadjuvant chemotherapy in patients with TNBC

BackgroundTriple-negative breast cancer (TNBC) is the most aggressive form of breast cancer (BC). Due to the absence of targets such as HER2 or hormone receptors, early TNBC is treated with...

Polymer-Lipid Hybrid Systems: Scope of Intravenous-To-Oral Switch in Cancer Chemotherapy

Cancer chemotherapeutic administration by oral route has the potential to create “hospitalization free chemotherapy”. Such a therapeutic approach will improve patient compliance and significantly reduce the cost of treatment. In current clinical practice, chemotherapy is primarily carried out by intravenous injection or infusion and leads to various unwanted effects. Despite the presence of oral delivery challenges like poor aqueous solubility, low permeability, drug stability and substrate for multidrug efflux transporter, cancer chemotherapy delivery through oral administration has gained much attention recently due to having more patient compliance compared to the intravenous mode of administration. In order to address the multifaceted oral drug delivery challenges, a hybrid delivery system is conceptualized to merge the benefits of both polymeric and lipid-based drug carriers. Polymer-lipid hybrid systems have presented various significant benefits as an efficient carrier to facilitate oral drug delivery by surmounting the different associated obstacles. This carrier system has been found suitable to overcome the numerous oral absorption hindrances and facilitate the intravenous-to-oral switch in cancer chemotherapy. In this review, we aimed to discuss the different biopharmaceutic challenges in oral delivery of cancer chemotherapy and how this hybrid system may provide solutions to such challenges.

Impact of chemotherapy-induced enteric nervous system toxicity on gastrointestinal mucositis.

Chemotherapy is a first-line treatment for many cancers; however, its use is hampered by a long list of side-effects. Gastrointestinal mucositis is a common and debilitating side-effect of anticancer therapy contributing to dose reductions, delays and cessation of treatment, greatly impacting clinical outcomes. The underlying pathophysiology of gastrointestinal mucositis is complex and likely involves several overlapping inflammatory, secretory and neural mechanisms, yet research investigating the role of innervation in gastrointestinal mucositis is scarce. This review provides an overview of the current literature surrounding chemotherapy-induced enteric neurotoxicity and discusses its implications on gastrointestinal mucositis. Damage to the intrinsic nervous system of the gastrointestinal tract, the enteric nervous system (ENS), occurs following chemotherapeutic administration, leading to altered gastrointestinal functions. Chemotherapeutic drugs have various mechanisms of actions on the ENS. Oxidative stress, direct toxicity and inflammation have been identified as mechanisms involved in chemotherapy-induced ENS damage. Enteric neuroprotection has proven to be beneficial to reduce gastrointestinal dysfunction in animal models of oxaliplatin-induced enteric neuropathy. Understanding of the ENS role in chemotherapy-induced mucositis requires further investigation and might lead to the development of more effective therapeutic interventions for prevention and treatment of chemotherapy-induced gastrointestinal side-effects.

Controlled delivery of a protein tyrosine phosphatase inhibitor, SHP099, using cyclodextrin-mediated host-guest interactions in polyelectrolyte multilayer films for cancer therapy.

The Src homology 2 domain containing protein tyrosine phosphatase-2 (SHP2) is a key enzyme in pathways regulating tumor growth signaling, and recently gained interest as a promising anticancer drug target. Many SHP2 inhibitors are currently under development, including SHP099, which has shown potent anticancer activity at low concentrations in vivo. In this work, we developed multilayer coatings for localized delivery of SHP099 to improve upon current cancer therapies. Layer-by-layer self-assembly was used to develop films composed of chitosan and poly-carboxymethyl-β-cyclodextrin (PβCD) for the delivery of SHP099. SHP099 was successfully loaded into multilayer films via host–guest interactions with PβCD. Nuclear magnetic resonance spectroscopy confirmed the occurrence of this supramolecular assembly by identifying the interaction of specific terminal SHP099 protons with the protons of the CD. SHP099 release from assembled films was detected over 96 hours, and was found to inhibit colony formation of human breast adenocarcinoma cells in vitro. These multilayer films have the potential to be used in a range of anticancer applications and overcome common complications of systemic chemotherapeutic administration, while maximizing SHP099 efficacy.

Abstract 3952: Localized nanodelivery combined with immunotherapy promotes curative anti-tumor responses in a murine breast cancer model

Abstract Optimal pairing of chemotherapy with immunotherapy can harness potential synergies and augment tumor immunity in solid tumors with low mutation burden by i) reducing tumor bulk, ii) releasing tumor antigen for cross-presentation and cross-priming, iii) releasing cancer-suppressive cytokines/chemokines, and iv) retaining circulating immune cell populations. Temperature-sensitive liposomes (TSL) have the potential to accomplish these goals as ultrasound (US) insonation or other treatment modalities can release the chemotherapy within bulky solid tumors in response to hyperthermia to maximize the local response and minimize off-target effects. Here we used Doxorubicin (Dox), stabilized within the core of TSL by forming a drug complex with copper (CuDox), to mediate immunogenic cell death (ICD) and release of type I interferons (IFNs). This activatable chemotherapy (CuDox-TSL+US) was combined with immunotherapy using CpG-ODN, a toll-like receptor 9 agonist, and a PD-1 antibody (αPD-1) for checkpoint blockade. We assessed the schedule and sequence of the chemo-immunotherapy protocol to maximize therapeutic outcome. CuDox-TSL were made of DPPC:MPPC:DSPE-PEG2k, 86:10:4 in the presence of copper (II) gluconate and triethanolamine at 0.2 mg-Dox/mg-lipid. Mice with bilateral invasive neu deletion (NDL) tumors (4 mm) were treated with an i.v. administration of CuDox-TSL at 6 mg Dox/kg body weight. One tumor was insonified with a peak ultrasound pressure of 1.1 MPa at a frequency of 1.5 MHz at 42°C for 5 min prior to and 20 min post drug injection with a variable duty cycle. Upon completion of US-hyperthermia, 100 µg of CpG-ODN 1826 was injected intratumorally to the insonified tumor. Three days later αPD-1 (200 µg, i.p.) was administrated. We found that Dox in combination with hyperthermia induced release of ICD markers and type I IFNs in cell culture of various murine cancer cell lines. Following intravenous administration of CuDox-TSL, ultrasonic activation of liposomes in the treated tumors promoted release and cross-presentation of a model tumor antigen by antigen-presenting cells in distant tumors. While a variety of chemo-immunotherapy protocols achieved a complete response in more than 50% of treated mice, the complete response rate was greatest (90%) when one week of immunotherapy priming preceded activatable chemotherapeutic administration. Repeated chemotherapeutic delivery, while effectively reducing viable tumor, also reduced efficacy and the fraction of viable CD8+ T-cells as indicated by flow cytometric analysis. Taken together, the results suggest that a single-dose administration of activatable chemotherapy after immune priming enhances survival in a murine model of breast cancer. In clinical practice, however, assessment of macrophage and T-cell populations over the course of treatment could help inform dose timing and number of treatment repetitions. Citation Format: Azadeh Kheirolomoom, Matthew T. Silvestrini, Elizabeth S. Ingham, Lisa M. Mahakian, Sarah M. Tam, Spencer K. Tumbale, Josquin Foiret, Neil E. Hubbard, Alexander D. Borowsky, William J. Murphy, Katherine W. Ferrara. Localized nanodelivery combined with immunotherapy promotes curative anti-tumor responses in a murine breast cancer model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3952.

Combining activatable nanodelivery with immunotherapy in a murine breast cancer model

A successful chemotherapy-immunotherapy solid-tumor protocol should accomplish the following goals: debulk large tumors, release tumor antigen for cross-presentation and cross-priming, release cancer-suppressive cytokines and enhance anti-tumor immune cell populations. Thermally-activated drug delivery particles have the potential to synergize with immunotherapeutics to accomplish these goals; activation can release chemotherapy within bulky solid tumors and can enhance response when combined with immunotherapy. We set out to determine whether a single protocol, combining locally-activated chemotherapy and agonist immunotherapy, could accomplish these goals and yield a potentially translational therapy. For effective delivery of free doxorubicin to tumors with minimal toxicity, we stabilized doxorubicin with copper in temperature-sensitive liposomes that rapidly release free drug in the vasculature of cancer lesions upon exposure to ultrasound-mediated hyperthermia. We found that in vitro exposure of tumor cells to hyperthermia and doxorubicin resulted in immunogenic cell death and the local release of type I interferons across murine cancer cell lines. Following intravenous injection, local activation of the liposomes within a single tumor released doxorubicin and enhanced cross-presentation of a model antigen at distant tumor sites. While a variety of protocols achieved a complete response in >50% of treated mice, the complete response rate was greatest (90%) when 1 week of immunotherapy priming preceded a single activatable chemotherapeutic administration. While repeated chemotherapeutic delivery reduced local viable tumor, the complete response rate and a subset of tumor immune cells were also reduced. Taken together, the results suggest that activatable chemotherapy can enhance adjuvant immunotherapy; however, in a murine model the systemic adaptive immune response was greatest with a single administration of chemotherapy.

Impact of Different Prognostic Factors on the Survival of Patients with Malignant Granulosa Cell Tumors of the Ovary: An Institutional Based Study at the Egyptian National Cancer Institute

Introduction: Emphasizing and evaluating different prognostic factors including the age, size of the tumor, type of surgery, capsular infiltration and administration of adjuvant chemotherapy, which may affect the disease free survival (DFS) and overall survival (OS) in patients with malignant Granulosa cell tumors (MGCTs). Patients and Methods: A retrospective study of 36 female patients diagnosed with MGCTs and managed at the National Cancer Institute – Cairo University-Egypt, in the period from May 2007 –October 2012. Follow –up reached up to 116 months (3-116 mo.) with a median of 72 months. Prognostic factors as the age, size of the tumor, type of surgery, capsular infiltration and administration of adjuvant chemotherapy with number of cycles in correlation with DFS and OS were statistically analyzed to detect any significant correlation. Results: The median age was 44 years. The median tumor size was 12 cm. The median DFS was 41.9 months. The median OS was 45.0 months. The disease free survival (DFS) was not significantly affected by the size of the tumor (<10 cm/ ≥10 cm) nor the type of surgery (Panhysterectomy or salpingo-oophorectomy) nor administration of adjuvant chemotherapeutic agents where p-value=0.4487, 0.8471 and 0.0986 respectively. However, capsular invasion significantly affected the DFS (p-value=0.0076). The overall survival (OS) was not affected by the size of the tumor (<10 cm/ ≥10 cm) with a nearly significant correlation (p=0.0615). No statistical significance on OS regarding the type of surgery (Panhysterectomy or salpingo-oophorectomy) (p=0.7792), the capsular affection (p=0.0871), nor the adjuvant chemotherapeutic administration (p= 0.2848). Conclusion: Capsular infiltration is by far the most independent prognostic factor affecting disease free survival (DFS) in patients with malignant granulosa cell tumors and to a lesser extent, overall survival (OS), Tumor size ≥10 cm is nearly affecting OS but not DFS. A Multi-institutional study with larger sample size and longer follow-up is needed to re-evaluate our results.

Doxorubicin Temporally Modulates Cyclooxygenase‐2 Levels in Male and Female Human Vascular Smooth Muscle Cells

Doxorubicin (Dox) is a chemotherapeutic agent that is highly effective at reducing recurrence and mortality in breast cancer patients. However, this anti‐cancer drug elicits dose‐ and time‐dependent cardiovascular toxicity. The pathogenesis of Dox is multifactorial, one of which involves the development and progression of inflammation mediated by activation of the TLR4/NFκB pathway. In a recent collaborative study, we demonstrated that intermittent bouts of Dox administered to mimic chemotherapeutic administration clinically attenuated protein levels of cyclooxygenase‐2 (COX‐2), a downstream proinflammatory mediator of the TLR4/NFκB pathway, in cerebral and peripheral vasculatures isolated from female ovariectomized rats. These findings were surprising and not in agreement with previously published data. Therefore, to address whether this Dox‐induced attenuated inflammatory response is time and dose dependent, we exposed cultured primary human female and male vascular smooth muscle cells to vehicle (saline) or Dox (0.3, 1.0, and 5.0 μM) for 6 and 24 hr. Following Dox treatment, cells were isolated, homogenized, and whole cell lysates analyzed for COX‐2 protein levels via immunoblotting. We also assessed levels of the upstream transcription factor, NFκBp65 as well as its activation. We observed that Dox dose dependently decreased levels of COX‐2 without altering NFκBp65 whole cell lysate protein levels at the 6 and 24 hr time points. However, NFκBp65 activity (6 hr time point), assessed indirectly using nuclear/cytosolic fractionation, was increased. Dox had no effect on cell morphology or density determined by light microscopy. Live versus dead cell percentages, determined using trypan blue, were also not affected by Dox suggesting that the anticancer drug did not alter cell death at 6 and 24 hr time points. In conclusion, these studies demonstrate a possible novel action for the anticancer agent eliciting both a dose‐ and time‐dependent effect on selective downstream mediators of inflammation such as COX‐2 in the female and male vasculature. Future studies are planned to evaluate the impact of acute versus chronic Dox treatment on COX‐2 at the transcriptional or translational levels to determine a mode of molecular regulation and to assess if sex differences exist.Support or Funding InformationUniversity of Arizona Sarver Heart Center Medical Student Award (MS) and the Valley Research Partnership P1 Grant (RJG).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.