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e24106Background: Circulating tumor cells (CTCs) have been demonstrated as a biomarker in certain advanced cancers. Additionally, there has been increasing recognition that definitive treatment, in...

Aim: The development of chemoradiation – the concurrent administration of chemotherapy and radiotherapy – has led to significant improvements in local tumor control and survival. However, it is limited by its high toxicity. In this study, we report the development of a novel NP (nanoparticle) therapeutic, ChemoRad NP, which can deliver biologically targeted chemoradiation. Method: A biodegradable and biocompatible lipid–polymer hybrid NP that is capable of delivering both chemotherapy and radiotherapy was formulated. Results: Using docetaxel, indium111 and yttrium90 as model drugs, we demonstrated that the ChemoRad NP can encapsulate chemotherapeutics (up to 9% of NP weight) and radiotherapeutics (100 mCi of radioisotope per gram of NP) efficiently and deliver both effectively. Using prostate cancer as a disease model, we demonstrated the targeted delivery of ChemoRad NPs and the higher therapeutic efficacy of ChemoRad NPs. Conclusion: We believe that the ChemoRad NP represents a new class of therapeutics that holds great potential to improve cancer treatment.

Nanotechnology, the area of study involving nanoparticles, century; medical fields. The nanotechnology is one of the fastest growing sectors of the high-tech economy in the 21st industry, it is currently undergoing unprecedented development in nowadays especially in areas of biological research for clinical, environmental and life science applications. The most frequently used nanoparticles are superparamagnetic iron oxide (SPIO) nanoparticles and semiconductor quantum dots (QDs). SPIO nanoparticles with nanometer sizes and superparamagnetic properties are widely used for labeling and sorting cells or organelles, magnetic resonance imaging, targeted drug delivery, and hyperthermia. QDs combined the unique size-dependent physical properties, excellent chemical and photochemical stability, controlled and increased endocytosis, enhanced cooperative binding activity, and easy introduction of multi-functionality for targeted delivery and imaging, and became the most promising labeling tool for life science studies. Both types of nanomaterials provide special approaches for complex studies and play very important roles in modern biomedical research. However, non-specific uptake of QDs and SPIO nanoparticles is a major concern because it can lead to false positives or false results. Hence, we report cellular uptake studies on SPIO nanoparticles and QDs conducted on breast cancer cell line SK-BR3. The results show that both QDs and SPIO nanoparticles have very strong non-specific cellular uptake. With specially designed blocking buffer (BBB) the non-specific signals were significantly decreased.

We report the development of early stage detection of breast cancer cells using quantum dots (QDs) as fluorescent signal generator for detection. The QDs based imaging of breast cancer cells involved two kinds of antibodies: the first was for labeling the cells and the second was for imaging the breast cancer cells. Anti-Her2/neu (10Ab) was used for labeling the captured SK-BR3 cells. The second antibody against anti-Her2/neu (20Ab) that was conjugated to QDs (20Ab~QDs) forms the complete assay, SK-BR3 + 10Ab + 20Ab~QDs, to generate the fluorescent cells for imaging. Fluorescent images of the complete assay for SK-BR3 cells were evaluated under a microscope with a UV light source. The preliminary results showed that the breast cancer cell SK-BR3 in the complete assay were successfully observed as fluorescent cells that had brighter signals compared with those labeled with organic dye using similar parameters and the same number of cells.

ThemajorshortcomingofCombidexisitsinabilitytodetectPCadiseaseoutsideofthelymphnodes.Herein, we report the development of a novel, multifunc-tional, thermally cross-linked SPION (TCL-SPION) that can bothdetect PCa cells, and deliver targeted chemotherapeuticagents directly to the PCa cells. We previously reported theuseoftheA10RNAaptamer (Apt), which bindstheextracellu-lar domain of the prostate-specific membrane antigen (PSMA),to engineer targeted nanoparticles for PCa therapy and imag-ing.

Oncogenic tyrosine kinases, such as BCR-ABL, TEL-ABL, TEL-PDGFβR, and FLT3-ITD, play a major role in the development of hematopoietic malignancy. They activate many of the same signal transduction pathways. To identify the critical target genes required for transformation in hematopoietic cells, we used a comparative gene expression strategy in which selective small molecules were applied to 32Dcl3 cells that had been transformed to factor-independent growth by these respective oncogenic alleles. We identified inhibitor of DNA binding 1 (Id1), a gene involved in development, cell cycle, and tumorigenesis, as a common target of these oncogenic kinases. These findings were prospectively confirmed in cell lines and primary bone marrow cells engineered to express the respective tyrosine kinase alleles and were also confirmed in vivo in murine models of disease. Moreover, human AML cell lines Molm-14 and K562, which express the FLT3-ITD and BCR-ABL tyrosine kinases, respectively, showed high levels of Id1 expression. Antisense and siRNA based knockdown of Id1-inhibited growth of these cells associated with increased p27Kip1 expression and increased sensitivity to Trail-induced apoptosis. These findings indicate that Id1 is an important target of constitutively activated tyrosine kinases and may be a therapeutic target for leukemias associated with oncogenic tyrosine kinases.

Traditional cancer treatments rely on the use of cytotoxic agents, and the choice of regimen is based on the cancer’s anatomical origin and histology. Advances in cancer genomics and molecular profiling have shown that different cancers can be driven by the same mutations/signaling pathways, and

Collagen IV-targeting peptide-conjugated basement membrane-targeting nanoparticles are successfully engineered to identify early-stage blood vessel injury induced by high-dose radiotherapy.

Parenteral and oral routes have been the traditional methods of administering cytotoxic agents to cancer patients. Unfortunately, the maximum potential effect of these cytotoxic agents has been limited because of systemic toxicity and poor tumor perfusion. In an attempt to improve the efficacy of cytotoxic agents while mitigating their side effects, we have developed modalities for the localized iontophoretic delivery of cytotoxic agents. These iontophoretic devices were designed to be implanted proximal to the tumor with external control of power and drug flow. Three distinct orthotopic mouse models of cancer and a canine model were evaluated for device efficacy and toxicity. Orthotopic patient-derived pancreatic cancer xenografts treated biweekly with gemcitabine via the device for 7 weeks experienced a mean log2 fold change in tumor volume of –0.8 compared to a mean log2 fold change in tumor volume of 1.1 for intravenous (IV) gemcitabine, 3.0 for IV saline, and 2.6 for device saline groups. The weekly coadministration of systemic cisplatin therapy and transdermal device cisplatin therapy significantly increased tumor growth inhibition and doubled the survival in two aggressive orthotopic models of breast cancer. The addition of radiotherapy to this treatment further extended survival. Device delivery of gemcitabine in dogs resulted in more than 7-fold difference in local drug concentrations and 25-fold lower systemic drug levels than the IV treatment. Overall, these devices have potential paradigm shifting implications for the treatment of pancreatic, breast, and other solid tumors.

Chemosensitizers can improve the therapeutic index of chemotherapy and overcome treatment resistance. Successful translation of chemosensitizers depends on the development of strategies that can preferentially deliver chemosensitizers to tumors while avoiding normal tissue. We hypothesized that nanoparticle (NP) formulation of chemosensitizers can improve their delivery to tumors which can in turn improve their therapeutic index. To demonstrate the proof of principle of this approach, we engineered NP formulations of two chemosensitizers, the PI3-kindase inhibitor wortmanin (Wtmn) and the PARP inhibitor olaparib. NP Wtmn and NP olaparib were evaluated as chemosensitizers using lung cancer cells and breast cancer cells respectively. We found Wtmn to be an efficient chemosensitizer in all tested lung-cancer cell lines reducing tumor cell growth between 20 and 60% compared to drug alone. NP formulation did not decrease its efficacy in vitro. Olaparib showed less consistent chemosensitization as a free drug or in NP formulation. NP Wtmn was further evaluated as a chemosensitizer using mouse models of lung cancer. We found that NP Wtmn is an effective chemosensitizer and more effective than free Wtmn showing a 32% reduction in tumor growth compared to free Wtmn when given with etoposide. Importantly, NP Wtmn was able to sensitize the multi-drug resistant H69AR cells to etoposide. Additionally, the combination of NP Wtmn and etoposide chemotherapy did not significantly increase toxicity. The present study demonstrates the proof of principle of using NP formulation of chemosensitizing drugs to improve the therapeutic index of chemotherapy.

Background This pilot trial is designed to determine whether PET/CT-guided radiotherapy dose escalation can improve local control while minimizing toxicity for the treatment of locally advanced nasopharyngeal carcinoma. Methods 67 patients were randomized into the three treatment arms: conventional chemoradiotherapy (group A), CT-guided dose escalation chemoradiotherapy (group B) and PET/CT-guided dose escalation chemoradiotherapy (group C). Radiotherapy was delivered using the simultaneous modulated accelerated radiation therapy (SMART) technique in the dose-escalation treatment arms. Patients received concurrent and adjuvant chemotherapy. Results The use of PET/CT significantly changed the treatment volume delineation of the gross tumor volume. 3-year local progression-free (LPF) survival rates of three groups were 83.3%, 90.9% and 100%, respectively. The 3-year regional progression-free survival (RPFS) rates were 95.8%, 95.5% and 100%, respectively. The 3-year disease free survival (DFS) rates were 79.2%, 86.4% and 95.2%, respectively. The 3-year overall survival (OS) rates were 83.3%, 90.9% and 95.2%, respectively. The 3-year disease-free survival (DFS) rates were 79.2%, 86.4% and 95.2%, respectively. No patient had grade 4 late toxicity. Conclusions PET/CT-guided dose escalation radiotherapy is well-tolerated and appears to be superior to conventional chemoradiotherapy for locally advanced NPC. Trial Registration ClinicalTrials.gov NCT02089204

Combining molecularly targeted agents and chemotherapeutics is an emerging strategy in cancer treatment. We engineered sub-50 nm diameter diblock copolymer nanoparticles (NPs) that can sequentially release wortmannin (Wtmn, a cell signaling inhibitor) and docetaxel (Dtxl, genotoxic anticancer agent) to cancer cells. These NPs were studied in chemoradiotherapy, an important cancer treatment paradigm, in the preclinical setting. We demonstrated that Wtmn enhanced the therapeutic efficacy of Dtxl and increased the efficiency of radiotherapy (XRT) in H460 lung cancer and PC3 prostate cells in culture. Importantly, we showed that NPs containing both Wtmn and Dtxl release the drugs in a desirable sequential fashion to maximize therapeutic efficacy in comparison to administering each drug alone. An in vivo toxicity study in a murine model validated that NPs containing both Dtxl and Wtmn do not have a high toxicity profile. Lastly, we demonstrated that Dtxl/Wtmn-coencapsulated NPs are more efficient than each single-drug-loaded NPs or a combination of both single-drug-loaded NPs in chemoradiotherapy using xenograft models. Histopathological studies and correlative studies support that the improved therapeutic efficacy is through changes in signaling pathways and increased tumor cell apoptosis. Our findings suggest that our nanoparticle system led to a dynamic rewiring of cellular apoptotic pathways and thus improve the therapeutic efficiency.

Purpose: Circulating endothelial cells (CEC) comprise at least two distinct populations: bone marrow–derived circulating endothelial progenitors (CEP) and mature CECs derived from existing vasculature. We hypothesized that antiangiogenic agents may have differential effects on CEPs and mature CECs and that these changes may serve as a marker of biological activity. Experimental Design: The effect of angiogenesis inhibitors on CECs was evaluated by flow cytometry after vascular endothelial growth factor (VEGF)–induced mobilization and in mice bearing Lewis lung carcinoma (LLC). Tumor angiogenesis was evaluated in parallel by immunohistochemistry. Results: In nontumor-bearing mice, VEGF administration increased both mature CECs and CEPs. This increase was inhibited by the VEGF receptor 2 inhibitor ZD6474 as well as the VEGF inhibitor–soluble Flt-1. ZD6474 had no significant effect on CECs in the absence of exogenous VEGF stimulation. In contrast, LLC-bearing mice had an increase in mature CECs but not CEPs after 3 days of treatment with ZD6474. The increase in mature CECs was dose-dependent, accompanied by a decrease in tumor microvessel density, and preceded reduction in tumor volume. Treatment of LLC-bearing mice with the vascular targeting agent ZD6126 also increased mature CECs. Conclusions: VEGF inhibitors can have differential effects on mature CECs and CEPs, and agents inhibiting tumor angiogenesis may cause a concomitant increase in mature CECs. This increase occurs in tumor-bearing but not in nontumor-bearing mice, suggesting that tumor endothelium is a potential source of mature CECs. Therefore, assessing both mature CECs and CEPs may provide insights into the mechanism of antiangiogenic agents and serve as an early surrogate marker of biological activity.

To test the hypothesis that HLA-B27 predisposes to disease by forming disulfide-linked homodimers, we examined rats transgenic for HLA-B27, mutant Cys67Ser HLA-B27, or HLA-B7. In splenic Con A blasts from high transgene copy B27 lines that develop inflammatory disease, the anti-H chain mAb HC10 precipitated four bands of molecular mass 78–105 kDa and additional higher molecular mass material, seen by nonreducing SDS-PAGE. Upon reduction, all except one 78-kDa band resolved to 44 kDa, the size of the H chain monomer. The 78-kDa band was found to be BiP/Grp78, and the other high molecular mass material was identified as B27 H chain. Analysis of a disease-resistant low copy B27 line showed qualitatively similar high molecular mass bands that were less abundant relative to H chain monomer. Disease-prone rats with a Cys67Ser B27 mutant showed B27 H chain bands at 95 and 115 kDa and a BiP band at 78 kDa, whereas only scant high molecular mass bands were found in cells from control HLA-B7 rats. 125I-surface labeled B27 oligomers were immunoprecipitated with HC10, but not with a mAb to folded B27-β2-microglobulin-peptide complexes. Immunoprecipitation of BiP with anti-BiP Abs coprecipitated B27 H chain multimers. Folding and maturation of B27 were slow compared with B7. These data indicate that disulfide-linked intracellular H chain complexes are more prone to form and bind BiP in disease-prone wild-type B27 and B27-C67S rats than in disease-resistant HLA-B7 rats. The data support the hypothesis that accumulation of misfolded B27 participates in the pathogenesis of B27-associated disease.

Purpose: To prospectively evaluate changes in circulating tumor cells (CTCs) following definitive treatment in patients with oligometastatic solid tumors using a novel nanotechnology-based biomimetric platform. Experiment Procedures: Patients with biopsy proven oligometastatic disease with up to three lesions were eligible if all sites of disease were treated with definitive therapy. Definitive therapy could include fractionated 3D conformal radiation, SBRT, IORT, surgical metastatectomy, and ablative procedures including microwave ablation. Patients initially presenting with oligometastatic disease were eligible if they had less than 3 lesions including the primary tumor. At least one lesion had to be treated with radiation. CTCs were enumerated from whole blood using the Onco-Sense CTC capture system which utilizes E-selectin and dendrimers functionalized with three cancer-specific antibodies (aEpCAM, aHER-2, aEGFR). Whole blood was collected from patients prior to starting treatment (baseline), mid-treatment, end of treatment, within 12 weeks of finishing treatment, and then every 3 months in follow up. We then assessed changes in CTCs with treatment and looked for associations between baseline CTCs, changes during treatment, and post-treatment changes with clinical outcomes. Results: We have currently enrolled 24 patients. The majority of patients (20) had a single site of disease treated with radiosurgery. Sites of metastasis include bone, lung, and lymph nodes. Primary histologies include lung, breast, melanoma, prostate, RCC, colorectal cancer, and SCC of the head and neck. CTCs were detected in 100% of patients at baseline and decreased with treatment from a mean of 45 CTCs/mL pretreatment (range 3-99) to 14 CTCs/mL post-treatment (range 3-59). Post-treatment CTCs are available for 14 patients with a median follow up of 10.3 months. Ten of 14 patients remain clinically NED and post-treatment CTCs are lower than baseline in 13/14 of these patients. There have been 4 clinical failures and CTCs increased with or before radiographic progression in all 4 patients. Three of the failures occurred at least 3 months after initial complete responses to treatment and CTCs increased by a median 3.4-fold over post-treatment levels in these patients. One patient progressed before the first post-treatment follow up and CTCs increased from 7 to 22 CTCs/mL from baseline to post-treatment. Conclusions: We showed here that we could enumerate CTCs in 100% of patients with oligometastatic disease using a novel CTC capture system. CTCs decreased with treatment and all clinical failures were preceded by significant rises in CTCs. Our preliminary data suggest that enumeration of CTCs by Onco-Sense may provide a novel biomarker for assessing treatment response and/or post-treatment surveillance for patients with oligometastatic solid tumors. Note: This abstract was not presented at the meeting. Citation Format: Joseph M. Caster, Kyle Wang, Bhisham Chera, Sin-Jun Park, Seungpyo Hong, Andrew Z. Wang. Assessing circulating tumor cells with a nanotechnology-based capture system as a novel biomarker for treatment response and surveillance in patients with oligometastatic solid tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1735. doi:10.1158/1538-7445.AM2017-1735

Background: Cancer immunotherapy is an exciting new approach to cancer treatment and there is strong interest in strategies to improve the long-term durable response rates of cancer immunotherapy. One approach is to combine checkpoint inhibitors such as aPD-1 with T cell activator such as OX40 agonist to further increase immune activation. We hypothesized that we can improve the therapeutic efficacy of this approach by temporally control the activation of OX40 and inhibition of PD-1 pathways. To accomplish this, we utilized nanoparticles that can deliver anti-OX40 and anti-PD1 antibodies simultaneously to T cells. Methods: Agonist antibody (anti-OX40) and antagonist antibody (anti-PD1) were conjugated to PLGA-PEG-Maleimide nanoparticles (AANPs) with precise ratio control and quantified by ELISA. Their specific binding to the target proteins was shown in vitro by flow cytometry. The tumor inhibition efficiency was assessed on mice bearing different tumor models. Two tumors were inoculated subcutaneously (105 B16F10 cells) or on fourth mammary fat pad (105 4T1 cells) on both flanks of mice. One side of tumor was irradiated once and AANPs were injected twice every 3 days. In vivo depletion experiments were tested on C56Bl6 mice and CD11b/c mice. Different populations of T cells in tumor and spleen were analyzed by flow cytometry and by fluorescent IHC staining. T cell killing assay and IFN-γ ELISpot were studied. Co-localization was demonstrated with fluorescent labeled antibodies and the corresponding AANPs. Results: AANPs showed a 30% cure rate, compared to 10% of free antibodies, 0% of anti-PD1 conjugated NPs, and 0% of anti-OX40 conjugated NPs in B16F10 melanoma model. We then re-challenged the cured mice with 2×105 B16F10 cells and none of the mice developed another tumor. In 4T1 breast cancer model, the survival rate on day 39 was 50% with AANPs treatment, compared to 22% in the mixture of anti-PD1 conjugated nanoparticles and anti-OX40 conjugated nanoparticles, and 0% of free antibodies. We demonstrated that AANPs led to a higher medium TCD8+/Treg ratio in tumors. The therapeutic effect was mediated by CD8+ T cells as elimination of these cells abrogated the therapeutic effects. In vitro study confirmed that AANPs were able to improve T cell stimulation compared to free antibodies by increasing IFN-γ excretion (2x). We further confirmed co-localization of antibodies with AANPs on tumor infiltration T cells in vivo. Our data demonstrated that spatial-temporal delivery of agonist and antagonist could improve T cell activation and cancer immunotherapy. Conclusions: Our data demonstrates that spatial-temporal delivery of agonist and antagonist can improve T cell activation and cancer immunotherapy. Citation Format: Yu Mi, Christof C. Smith, Feifei Yang, Jonathan Serody, Benjamin Vincent, Andrew Z. Wang. Spatial-temporal delivery of OX40 agonist and PD-1 inhibitor using nanoparticles improves therapeutic efficacy of cancer immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 978. doi:10.1158/1538-7445.AM2017-978

11533 Background: Circulating tumor cells (CTCs) can provide prognostic information in select patients with advanced cancers. We have developed a sensitive and specific CTC capture system (Oncosensor) which may predict clinical outcomes in patients undergoing definitive radiation for head and neck cancers. There is increasing interest in utilizing potentially curative metastasis-directed therapy for patients with oligometastatic solid-tumor malignancies. There is currently no biomarker to predict the success of this approach in most patients. The purpose of this prospective study is to investigate the potential utility of CTCs as a predictive biomarker for patients with oligometastatic cancer undergoing potentially curative therapy. Methods: Eligible patients had a metastatic solid-tumor malignancy with 3 or fewer metastases. All sites of disease were treated with definitive radiation or surgery. Concurrent chemotherapy was allowed. Peripheral blood (7 ml) was collected prior to starting, first week, mid-point, end RT, and every 4 to 12 weeks post RT. CTCs were quantified using the Oncosensor platform. We then assessed correlations between CTCs (baseline, end treatment, and changes during treatment) and clinical outcomes using multivariate analysis. Results: Baseline CTCs were detected in 20/20 enrolled patients with a mean baseline of 32/ml which decreased to a mean of 14/ml at the end of treatment. There was no association between pretreatment CTCs and clinical outcomes (p = 0.81). There was a significant association between post-RT CTCs and PFS (p = 0.039, HR 1.07 per CTC). Our data also suggest that post-treatment CTC monitoring may be able to detect early disease recurrence. Among the 4 patients with documented clinical failures and post-treatment CTC monitoring, all 4 had increases in CTCs with or prior to clinical or radiographic disease progression, with 1.6 to 7.3 fold increase at the time of progression compared to the prior time point. Conclusions: Our pilot data suggest CTCs may provide a predictive biomarker for patients with oligometastatic disease and could potentially provide a novel marker of disease recurrence.

Nanotechnology, the manipulation of matter on atomic and molecular scales, is a relatively new branch of science. It has already made a significant impact on clinical medicine, especially in oncology. Nanomaterial has several characteristics that are ideal for oncology applications, including preferential accumulation in tumors, low distribution in normal tissues, biodistribution, pharmacokinetics, and clearance, that differ from those of small molecules. Because these properties are also well suited for applications in radiation oncology, nanomaterials have been used in many different areas of radiation oncology for imaging and treatment planning, as well as for radiosensitization to improve the therapeutic ratio. In this article, we review the unique properties of nanomaterials that are favorable for oncology applications and examine the various applications of nanotechnology in radiation oncology. We also discuss the future directions of nanotechnology within the context of radiation oncology.

Aptamers, single-stranded oligonucleotides, are an important class of molecular targeting ligand. Since their discovery, aptamers have been rapidly translated into clinical practice. They have been approved as therapeutics and molecular diagnostics. Aptamers also possess several properties that make them uniquely suited to molecular imaging. This review aims to provide an overview of aptamers’ advantages as targeting ligands and their application in molecular imaging.

The effects of nanoparticle (NP) properties, such as size, shape and surface charge, on their efficacy and toxicity have been studied extensively. However, the effect of controlled drug release on NP efficacy and toxicity has not been thoroughly evaluated in vivo. Our study aims to fill this knowledge gap. A key challenge in characterizing the relationship between drug release and therapeutic ratio is to fabricate NPs that differ only in their drug release profile but are otherwise identical. To overcome this challenge, we developed crosslinkable lipid shell (CLS) NPs, where the drug release kinetics can be modulated without changing any other NP property. Using CLS NPs with wortmannin and docetaxel as model drugs, we determined the relationship between the release kinetics and therapeutic efficacy and toxicity of the drugs. We have determined that drug release kinetics can affect the therapeutic efficacy of NP docetaxel and NP wortmannin in vitro and in vivo. Our study also demonstrates that a decrease in drug release kinetics can result in a decrease in the hepatotoxicity of CLS NP wortmannin. Using two model drugs, the current findings provide the first direct evidence that NP drug release profile is a critical factor in determining the NP therapeutics' efficacy and toxicity in vivo.

Nanoparticles can be engineered with distinctive compositions, sizes, shapes, and surface chemistries to enable novel techniques in a wide range of biological applications. The unique properties of nanoparticles and their behavior in biological milieu also enable exciting and integrative approaches to studying fundamental biological questions. This review will provide an overview of various types of nanoparticles and concepts of targeting nanoparticles. We will also discuss the advantages and recent applications of using nanoparticles as tools for drug delivery, imaging, sensing, and for the understanding of basic biological processes.

Current preclinical evaluations of nanoparticle taxanes have focused on the effect of nanoparticle size and shape on the efficacy and toxicity. It is generally assumed that nanoparticle therapeutics have the same cellular response on tumor and normal cells as their small molecule counterparts. Here, we show that nanoparticle taxanes can mediate cellular effects distinct from that of small molecule taxanes at the sub-therapeutic dose range. Cells that are exposed to two polymeric nanoparticle formulations of docetaxel were found to undergo a different cell cycle and cell fate than those of cells that were exposed to small molecule docetaxel. Our results suggest that nanoparticle formulation of therapeutics can affect the therapeutic effect of its cargo. From the Clinical Editor This study investigates the differences between subtherapeutic doses of docetaxel applied as small molecules vs. nanoparticle formulations, demonstrating differential effects on the cell cycle and overall cell fate. The study suggests that the carrier may change the therapeutic effects of its cargo, which has important implications on future research.

Quantum dots (QD) nanoparticles have been widely used in biomedical and electronics fields, because of their novel optical properties. Consequently it confers enormous potential for human exposure and environmental release. To increase the biocompatibility of QDs, a variety of surface coatings or functional groups are added to increase their bioactivity and water solubility. Human adult low calcium high temperature (HaCaT) cells are the epithelial cells derived from adult human skin that exhibits normal differentiation capacity and a DNA fingerprint pattern that is unaffected by long-term cultivation, transformation, or the presence of multiple chromosomal alternations. Human keratinocytes, HaCaT cells were used to systematically evaluate the cytotoxicity of biocompatible QD made of CdSe metal core and ZnS shell with three different coatings and at three different wavelengths (530, 580 and 620 nm). In terms of half-maximal inhibitory concentration, QSA-QDs with amine-polyethyleneglycol coating and QSH-QDs with amphiphilic polymer coating were not cytotoxic, while QEI-QDs with polyethylenimine coating were highly toxic to the HaCaT cells in comparison to a reference CuInS2/ZnS. QEI-QDs led to significant increase in reactive oxygen species, decrease in mitochondrial membrane potential and DNA damage in HaCaT cells. The mechanisms of toxicity of QEI-530 and QEI-580 can be attributed to the combination of intracellular reactive oxygen species production and loss of MMP. The QDs toxicity can be attributed to the polyethylemimine surface coating which was highly toxic to cells in comparison with amine-polyethyleneglycol, but not due to the release of cadmium ions.

Biotargeted nanomedicines have captured the attention of academic and industrial scientists who have been motivated by the theoretical possibilities of the ‘magic bullet’ that was first conceptualized by Paul Ehrlich at the beginning of the 20th century. The Biotargeting Working Group, consisting of more than 50 pharmaceutical scientists, engineers, biologists and clinicians, has been formed as part of the National Cancer Institute’s Alliance for Nanotechnology in Cancer to harness collective wisdom in order to tackle conceptual and practical challenges in developing biotargeted nanomedicines for cancer. In modern science and medicine, it is impossible for any individual to be an expert in every aspect of biology, chemistry, materials science, pharmaceutics, toxicology, chemical engineering, imaging, physiology, oncology and regulatory affairs. Drawing on the expertise of leaders from each of these disciplines, this commentary highlights six tenets of biotargeted cancer nanomedicines in order to enable the translation of basic science into clinical practice.

Prostate cancer treatment failure after local therapy can be detected through changes in PSA prior to any clinical evidence of disease. Therefore, posttreatment elevation, also called biochemical failure, has been utilized as a surrogate for disease recurrence. The ability to consistently define biochemical failure is important both for the prognostic risk stratification of patients for clinical recurrence and survival and for the standardization of research when comparing multiple series of patients who undergo the same treatment. Due to the difference in PSA posttreatment, the definition of biochemical failure varies between treatments. In this chapter, we will explore the definitions of biochemical failure for each type of prostate cancer treatment, the clinical workup at time of biochemical failure, and the management decisions that clinicians encounter after patients develop biochemical failure.

This study explored the novel use of iron oxide (IO) nanoparticles (90%) internalized IO nanoparticles, but only the latter were significantly activated, with elevated expression/secretion of CD86, cytokines (IL-6, TNF-α, IL1-b, IFN-γ, and IL-12), and chemokines (CXCL1, CXCL2, CCL2, CCL3, CCL4, and CXCL10). Thus, the IO nanoparticles is a novel, safe, and effective vaccine platform, with built-in adjuvancy, that is highly stable and field deployable for cost-effective vaccine delivery.—Pusic, K., Aguilar, Z., McLoughlin, J., Kobuch, S., Xu, H., Tsang, M., Wang, A., Hui, G. Iron oxide nanoparticles as a clinically acceptable delivery platform for a recombinant blood-stage human malaria vaccine.

Background: Circulating tumor cells (CTCs) are an important biomarker in cancer. There has been substantial interest in utilizing CTCs to enable personalized treatment. However, there is limited data on CTCs in patients with head and neck cancers (HNC) where there is growing data that curative treatment needs to be risk-based. The purpose of this prospective, correlative study is to investigate the clinical significance of CTCs, as measured by a highly sensitive CTC capture technology UiChip, in HNC patients undergoing definitive treatment. Methods: HNC patients (M0) undergoing definitive treatment (radiation+/- chemotherapy) were enrolled. Peripheral blood was collected prior to starting RT, at the first week, mid-point, final week of treatment, and every 4 to 12 weeks after completion. Quantification of CTCs was conducted using UiChip. The primary endpoint was change in CTCs pre- and post- RT. Results: 35 patients are included in this analysis (median age 58, 34% non-smokers, 69% HPV/p16 positive, and 17% node (-)) and 90% received chemotherapy. CTCs were detected in all patients pre-RT (100%). There was no association between pre-RT CTC level and tumor or nodal stage. Median CTCs significantly decreased from 71 CTCs/mL (range, 7-849) before RT to 32 (2-209) at completion of RT and 27 (2-78) at 4 to 12 weeks post-treatment. Ten patients had persistent disease and/or distant metastasis at a median follow-up of 10.6 months (range, 3.7-17). CTCs declined throughout treatment in patients with complete clinical and/or radiographic responses, in contrast to an elevation in CTCs at mid or post-RT in the 7 patients with pathologic residual disease/distant failures. Conclusions: We demonstrated that the novel UiChip can capture CTCs in a diverse population of head and neck cancer patients. Our pilot data suggest that individual patient CTC changes during and after treatment may be a predictive biomarker for radiotherapy response and clinical outcomes. Table: Median CTCs/mL Peripheral Blood (Median% of Pre-Treatment Baseline Count)Pre-RTMid-RTPost-RTNumber (%) of CTCs in Complete Responders to RT (N = 18)67 IQR: 39-11737 (38%) IQR: 22-79 (18-94%)21 (18%) IQR: 7-40 (13-42%)Number (%) of CTCs in Partial Responders to RT (N = 7)120 IQR: 36-30366 (123%) IQR: 20-134 (30-172%)51 (45%) IQR: 23-54 (17-63%) Citation Format: Joseph M. Caster, Michael J. Eblan, Kyle Wang, Ja Hye Myung, Bhishamjit Chera, Seungpyo Hong, Andrew Z. Wang. Prospective evaluation of circulating tumor cells (CTCs) in head and neck cancer patients receiving definitive radiotherapy with a nanotechnology based system. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3954.

Purpose: We sought to use combretastatin A-4 phosphate disodium (CA4P) to enhance the tumor uptake of doxorubicin (Dox)-loaded, polyethylene glycol (PEG)-coated hollow gold nanospheres (HAuNS; Dox@PEG-HAuNS) for improved photothermal ablation (PTA)-chemotherapy of hepatocellular carcinoma (HCC) in rats under image guidance. Materials and Methods: Two groups of 10 HCC-bearing rats each received intrahepatic arterial (IA) injection of either PEG-HAuNS/Lipiodol alone or CA4P followed by PEG-HAuNS/Lipiodol 5 min later. Each group was subdivided, and the Au content of tumors and tissues excised at 1 h or at 24 h was quantified using neutron activation analysis (n = 5/time point). Five rats received CA4P plus PEG-[64Cu]-HAuNS/Lipiodol and underwent μPET/CT. Therapeutic effects were compared among 3 groups of 6 rats each that received IA injection of saline (control group), CA4P plus Dox@PEG-HAuNS/Lipiodol (chemotherapy group), or CA4P plus Dox@PEG-HAuNS/Lipiodol plus near-infrared irradiation (PTA-chemotherapy group). Findings were verified with postmortem histopathology and/or autoradiography. Results: PEG-HAuNS uptake in CA4P-pretreated tumors was significantly higher than that in non-CA4P-pretreated tumors at both 1 h (P Citation Format: Junjie Li, Min Zhou, Chiyi Xiong, Wanqin Wang, Qizhen Cao, Xiaoxia Wen, Xin Ji, Andrew Wang, Sanjay Gupta, Chun Li. Intraarterial delivery of doxorubicin-loaded hollow gold nanospheres for photothermal ablation chemotherapy of hepatocellular carcinoma in the liver of rats. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4204.

Background: Metastasis contributes to majority of death in cancer patients. However, cancer metastasis is difficult to study due to the lack of experimental models that can fully recapitulate the biological process, especially the organ specificity of cancer metastasis. Recent advances in tissue engineering demonstrated that decellularized tissue, organs that are denuded of cells, are excellent scaffolds for tissue engineering of complex organs, such as liver and lung. These decellularized organs preserve the organ microenvironment, which is critical in cancer metastasis. We hypothesized that we can utilize decellularized organ and engineer in vitro models of cancer metastasis that can recapitulate the organ specificity of cancer metastasis. Here we report the proof-of-principle of this approach by employing colorectal cancer as a disease model. Since the primary cause of morbidity and mortality in colorectal patients is liver and lung metastasis, we aimed to engineer in vitro models of colorectal cancer lung and liver metastasis. Methods: Decellularized biomatrix was prepared by sodium deoxycholate based perfusion decellularization of rat liver or lung. We compared the growth factors on the biomatrix scaffold to that of normal rat liver and lung using semi-quantitative ELISA. We cultured colorectal cancer cell lines: HT-29, SW480 and Caco2, on tissue culture dishes coated with liver and lung decellularized biomatrix scaffolds. Cell colony morphology and structure was examined by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and hematoxylin and eosin stain. Therapeutic response (chemotherapy and radiation) for cells grown on biomatrix was also examined. Results: Decellularized organs preserved 92% growth factors bound to biomatrix scaffold at near physiological levels. HT-29, SW480 and Caco2 were able to proliferate on dishes coated with biomatrix scaffold and spontaneously formed three-dimensional (3D) colonies when maintained at higher levels. These tumor colonies are millimeter in size and are spherical in shape, similar to in vivo metastases. SEM showed cells on the tumor surface appeared to form tight junctions with each other. TEM images further confirmed that these 3D tumor colonies contain areas of necrosis, consistent existing literature 3D colonies and in vivo metastasis. Importantly, we identified signet ring cell formation in the engineered 3D metastases. We also found the cells in engineered liver and lung metastasis responded to chemotherapy treatment differently than the cells cultured under standard conditions or on collagen or on matrigel. Interestingly, cells in both engineered liver and lung metastasis are more sensitive to radiotherapy than standard condition. Conclusions: Our engineered in vitro tumor metastasis models closely mimic the in vivo metastasis phenotypically, histologically, and biologically. Citation Format: XI TIAN, Michael E. Werner, Henry P. Foote, Ariel D. Hanson, Lola M. Reid, Andrew Z. Wang. Engineered in vitro models of cancer metastasis using decellularized biomatrix. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4264.

Purpose: To compute daily dose delivered during radiotherapy, deformable registration needs to be relatively fast, automated, and accurate. The aim of this study was to evaluate the performance of commercial deformable registration software for deforming between two modalities: planning computed tomography (pCT) images acquired for treatment planning and cone beam (CB) CT images acquired prior to each fraction of prostate cancer radiotherapy. Methods: A workflow was designed using MIM Software™ that aligned and deformed pCT into daily CBCT images in two steps: (1) rigid shifts applied after daily CBCT imaging to align patient anatomy to the pCT and (2) normalized intensity-based deformable registration to account for interfractional anatomical variations. The physician-approved CTV and organ and risk (OAR) contours were deformed from the pCT to daily CBCT over the course of treatment. The same structures were delineated on each daily CBCT by a radiation oncologist. Dice similarity coefficient (DSC) mean and standard deviations were calculated to quantify the deformable registration quality for prostate, bladder, rectum and femoral heads. Results: To date, contour comparisons have been analyzed for 31 daily fractions of 2 of 10 of the cohort. Interim analysis shows that right and left femoral head contours demonstrate the highest agreement (DSC: 0.96±0.02) with physician contours. Additionally, deformed bladder (DSC: 0.81±0.09) and prostate (DSC: 0.80±0.07) have good agreement with physician-defined daily contours. Rectum contours have the highest variations (DSC: 0.66±0.10) between the deformed and physician-defined contours on daily CBCT imaging. Conclusion: For structures with relatively high contrast boundaries on CBCT, the MIM automated deformable registration provided accurate representations of the daily contours during treatment delivery. These findings will permit subsequent investigations to automate daily dose computation from CBCT. However, improved methods need to be investigated to improve deformable results for rectum contours.

4536Background: Trimodality BPT in MIBC includes a maximal transurethral resection followed by concurrent chemoradiotherapy as an alternative to cystectomy in appropriately selected patients, or as...

10102Background: Survivorship care planning upon treatment completion is especially important for prostate cancer patients treated with ADT. These patients need coordinated oncologic and primary ca...

11536Background: Circulating tumor cells (CTCs) are an important biomarker in cancer and there is significant interest in developing new technologies to adequately assess CTCs to enable personalize...

Study Type - Therapy (phase 1) Level of Evidence 2a What's known on the subject? and What does the study add? High-risk and locally advanced prostate cancers are difficult to cure with the standard regimen of radiation therapy (RT) with concurrent androgen-deprivation therapy (ADT). Multiple studies have explored the addition of docetaxel chemotherapy in attempt to improve patient outcomes. Prior Phase I studies have shown that docetaxel 20 mg/m(2) is a safe dose, when given concurrently with 70 Gy of radiation. But current standard RT for prostate cancer uses higher doses, and it is unclear if concurrent chemotherapy is safe with modern RT. This is a Phase I study that explored the addition of concurrent docetaxel chemotherapy to modern RT (intensity-modulated RT to 78 Gy) plus ADT. The study showed that weekly docetaxel at 20 mg/m(2) is safe with modern RT. At a median follow-up of 2.2 years, biochemical progression-free survival was 94%. This triple-therapy regimen is safe and promising for further evaluation in prospective trials.• To evaluate in a phase I trial, the feasibility of adding concurrent weekly docetaxel chemotherapy to high-dose intensity modulated radiation therapy (IMRT) and androgen-deprivation therapy (ADT) for treatment of high-risk prostate cancer.• Patients with high-risk prostate cancer were treated with a luteinising hormone-releasing hormone agonist (starting 2-3 months before IMRT and lasting 2 years), IMRT of 78 Gy to the prostate and seminal vesicles, and weekly docetaxel during RT. • All patients had computed tomography and bone scans to exclude metastatic disease. • A standard 3 + 3 design was used for docetaxel dose escalation. Successive patients were treated on dose levels of 10, 15, and 20 mg/m(2) of weekly docetaxel.• In all, 18 patients participated in the study: 15 (83%) had Gleason 8-10 disease; the other three had either clinical T3 disease and/or a prostate-specific antigen (PSA) level of20 ng/mL. • Grade 3 diarrhoea (a defined dose-limiting toxicity, DLT) occurred in one patient in each of the first two dose levels. However, when the cohorts were expanded, no further DLT was seen. • Weekly docetaxel at 20 mg/m(2) (dose level 3) was successfully given without DLT. • No patient had grade 4 or 5 toxicity. • At a median follow-up of 2.2 years, all patients achieved a PSA nadir of1 ng/mL, including 13 patients who had an undetectable PSA level. The 2-year biochemical progression-free survival was 94%.• A dose of 20 mg/m(2) of weekly docetaxel given concurrently with high-dose IMRT and ADT appears safe for further study in patients with high-risk prostate cancer.

One of the promises of nanoparticle (NP) carriers is the reformulation of promising therapeutics that have failed clinical development due to pharmacologic challenges. However, current nanomedicine research has been focused on the delivery of established and novel therapeutics. Here we demonstrate proof of the principle of using NPs to revive the clinical potential of abandoned compounds using wortmannin (Wtmn) as a model drug. Wtmn is a potent inhibitor of phosphatidylinositol 3′ kinase-related kinases but failed clinical translation due to drug-delivery challenges. We engineered a NP formulation of Wtmn and demonstrated that NP Wtmn has higher solubility and lower toxicity compared with Wtmn. To establish the clinical translation potential of NP Wtmn, we evaluated the therapeutic as a radiosensitizer in vitro and in vivo. NP Wtmn was found to be a potent radiosensitizer and was significantly more effective than the commonly used radiosensitizer cisplatin in vitro in three cancer cell lines. The mechanism of action of NP Wtmn radiosensitization was found to be through the inhibition of DNA-dependent protein kinase phosphorylation. Finally, NP Wtmn was shown to be an effective radiosensitizer in vivo using two murine xenograft models of cancer. Our results demonstrate that NP drug-delivery systems can promote the readoption of abandoned drugs such as Wtmn by overcoming drug-delivery challenges.

Pancreatic cancer is the fourth deadliest form of cancer with a dismal 5-year survival rate of less than 5%. As a result there is an urgent need for the development of more efficacious therapies for pancreatic cancer. In order to overcome the immune suppression that is prevalent in this disease, the use of immune checkpoint blockades, CTLA-4 and PD-L1, have been investigated. However, published reports have indicated that pancreatic cancer is generally non-responsive to these immunotherapies. Because it has been shown that combinatorial therapies consisting of immunotherapy and chemotherapy can act synergistically, we sought to investigate the efficacy of targeted delivery of the chemotherapeutic cisplatin using nanoparticle carriers along with PD-L1 blockade in a highly aggressive orthotopic model of pancreatic cancer derived from the PANC02 mouse cell line. Our lab has developed a receptor targeted magnetic iron oxide nanoparticle (IONP) platform for targeted tumor imaging and drug delivery and demonstrated its effect on tumor growth inhibition in breast, ovarian, and pancreatic cancer mouse models. Active targeting of IONPs requires conjugation of the recombinant amino terminal fragment (ATF) of the urokinase plasminogen activator (uPA) that will bind specifically to tumor cells and tumor associated stromal cells that express a high level of its receptor (uPAR). uPAR expression is associated with increased invasiveness and a poorer prognosis, which further makes this protein an ideal target. In the orthotopic PANC02 model, we found that intraperitoneal delivery of 100µg of PD-L1 blockade alone (clone 10F.9G2) twice per week for four injections reduced orthotopic tumor burden by ~30% relative to untreated mice. Whereas, the combination therapy with uPAR-targeted IONP-cisplatin (10mg/kg drug equivalent) decreased tumor burden by ~65%. Enhanced inhibition of ascites fluid accumulation was also observed following combinatorial therapy, 81% versus 55.6%, respectively. We observed that a high level of uPAR-targeted IONP-cisplatin accumulated in the orthotopic pancreatic tumor following intraperitoneal delivery by Prussian blue staining indicating the effective delivery of the chemotherapeutic drug to the tumor site which could mediate tumor growth inhibition. Additionally, this therapeutic synergy is likely due, in part, to the enhanced intratumoral infiltration of T cells following targeted IONP-drug treatment. Taken together, these findings indicate that the minimal therapeutic efficacy of immune checkpoint blockades such as PD-L1 in pancreatic cancer can be enhanced by the tumor-targeted delivery of chemotherapeutic drugs using theranostic nanoparticles and this combinatorial therapeutic regime warrants further investigation. Citation Format: Erica N. Bozeman, Ning Gao, Weiping Qian, Andrew Wang, Lily Yang. Synergistic effect of targeted chemotherapy delivery using theranostic nanoparticles and PD-L1 blockade in an orthotopic mouse pancreatic cancer model. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr A60.

Nanomedicine, the medical application of nanotechnology, is poised to make a major impact on the diagnosis and treatment of many diseases. It is a relatively new branch of science that involves harnessing the unique properties of particles that are nanometers in scale (nanoparticles) for both diagnostic and therapeutic applications. Nanoparticles (NPs) can be engineered with precise sizes, shapes, compositions and surface chemistries for specific applications. In particular, nanoparticles are uniquely suited for the treatment of cancers and cardiovascular diseases. NPs can be engineered with surface targeting ligands against cancer markers or atherosclerosis markers, which in turn allow the specific detection of as well as specific drug delivery to these diseases. In the case of cancer, nanoparticles also preferentially accumulate in tumors due to the enhanced permeability and retention (EPR) effect (Noguchi et al. 1998). Although nanomedicine is a relatively new field, it has been quickly translated into clinical medicine and has led to a number of clinically-approved agents. Today, there are more than twenty nanoparticle therapeutics in clinical use. Furthermore, a 2006 global survey conducted by the European Science and Technology Observatory (Duconge et al.) revealed that more than 150 companies are developing nanoscale therapeutics, highlighting the high enthusiasm for nanoparticle-based therapeutics (Wagner et al. 2006). While nanomedicine has many facets, radioisotopes have been an integral component. Radioisotopes have been extensively utilized in the development of nanoparticle-based therapeutics. Radiolabeling has been the predominant method for the accurate assessment of biodistribution, circulation half-life, and pharmacokinetics of nanoparticles. In addition, there has been great interest in developing nanoparticles with dual diagnostic and therapeutic functionalities (theranostics) and radioisotopes have been a natural fit for the diagnostic component. Lastly, therapeutic radioisotopes can also be delivered by nanoparticles, especially for the treatment of cancer. In this chapter, we will review the various applications of radioisotopes in nanomedicine in detail. We also plan to discuss the methods of incorporating radioisotopes into nanoparticles as well as preclinical and clinical data on radioisotope containing nanoparticle diagnostics and therapeutics.