Intratumoral Therapy Research Articles

Intratumoral Gene Therapy Injections with a Multipronged, Multi–Side Hole Needle for Rectal Carcinoma

Interventional radiologists can deliver therapeutics directly to a tumor via needle injection. One of the major limitations of directly injected gene therapies is limited delivery, penetration, and transfection efficiency of the genetic vector, which in many cases directly limits the therapeutic effect. Numerous preclinical oncology models exist which rely upon direct injection of DNA plasmid or vector through an end-hole needle and into a tumor. Selection of a physically more efficient device with more needles and more side holes could improve distribution of the therapeutic agent.[1] Small hepatocellular carcinomas have been effectively treated with alcohol injections via a multi-pronged, multi-side hole needle. [2] TNFerade (GenVec, Gaithersburg, MD) is an attenuated adenovirus vector whose cargo is the gene encoding for tumor necrosis factor-alpha (TNFα), a potent cytokine that can be tumorocidal and also makes nearby tumor cells more sensitive to chemotherapy and to radiation therapy. TNFerade is turned on by a promoter that is turned on by radiation. [3] TNFerade also causes a “bystander effect” whereby affected cells do not necessarily need to be transfected with virus, but can respond to the TNFα product programmed by the gene. Despite the bystander effect, injectable therapeutics may suffer from lack of exposure to tumor tissue or a limited distribution in target tumor. Optimizing the physical distribution of such a therapeutic agent should be a goal of the interventional radiologist performing direct gene therapy injections. 5 patients with T3, T4, or N+ rectal cancer were treated as part of a clinical trial with weekly transrectal ultrasound (TRUS) guided injections, monitored under direct TRUS visualization [fig 1]. Patients received injections via a Quadrafuse needle (Rex Medical, Conshohocken, PA & Alliance Medical, Austin, TX) [fig 1] with three prongs, each with three side holes, or a custom Bernardino needle (Cook Medical, Bloomington, IN) with three side holes. Five weekly injections of 4 × 1010 particle units TNF-erade were combined with 50.4 Gy of external beam radiation, chemotherapy with capecitibine, and surgical resection. No complications were seen from the 25 injection procedures in 5 patients. [3] Fig 1 Multi-prong, multi-side hole needle in rectal cancer monitoring gene therapy injection with ultrasound. Ultrasound shows central cannula (open arrow) has deployed prong (closed arrow) with side holes, to better distribute therapeutic agent. Direct injection of gene therapy is highly inefficient, which may limit transfection and theoretically limit therapeutic efficacy. Interventional radiologists should consider the speculative benefits of multi-pronged and multi-side hole needles when directly injecting gene therapies.

Abstract LB-129: CD8+ T cell response mediates the therapeutic effects of oncolytic adenovirus in an immunocompetent mouse model

Abstract The role of anti-tumor immune responses in oncolytic adenoviral therapy has not been well studied due to lack of efficacious tumor model in immunocompetent mice. Here, we evaluated the contributions of immune components to the therapeutic effects of oncolytic adenoviruse in an immunocompetent murine tumor model permissive for infection and replication of adenovirus. We found that CD8+ T cells were critical mediator for antitumor efficacy by oncolytic adenovirus. Intratumoral viral therapy induced intensive infiltration of CD8+ T cells in tumor, increased tumor-specific IFN-γ (interferon-γ) production and CTL (cytotoxic T lymphocyte) activity of lymphocytes, and generated a long-term tumor-specific immune memory. Boosting CD8+ T cell responses by agonistic anti-4-1BB (cluster differentiation 137, CD137) antibody showed synergistic anticancer effects with oncolytic viro-therapy. Our results provide insight into antitumor mechanisms of oncolytic adenovirus in addition to their direct oncolytic effect. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-129. doi:1538-7445.AM2012-LB-129

Intratumoral versus Intravenous Gene Therapy Using a Transcriptionally Targeted Viral Vector in an Orthotopic Hepatocellular Carcinoma Rat Model

PurposeTo evaluate the feasibility of intratumoral delivery of adenoviral vector carrying a bidirectional two-step transcriptional amplification (TSTA) system to amplify transcriptional strength of cancer-specific Survivin promoter in a hepatocellular carcinoma model. Materials and MethodsMCA-RH7777 cells were implanted in rat liver, and tumor formation was confirmed with [18F]fluorodeoxyglucose (18F-FDG) positron emission tomography (PET). The adenoviral vector studied had Survivin promoter driving a therapeutic gene (tumor necrosis factor-α–related apoptosis-inducing ligand [TRAIL]) and a reporter gene (firefly luciferase [FL]; Ad-pSurvivin-TSTA-TRAIL-FL). Tumor-bearing rats were administered Ad-pSurvivin-TSTA-TRAIL-FL intravenously (n = 7) or intratumorally (n = 8). For control groups, adenovirus FL under cytomegalovirus (CMV) promoter (Ad-pCMV-FL) was administered intravenously (n = 3) or intratumorally (n = 3). One day after delivery, bioluminescence imaging was performed to evaluate transduction. At 4 and 7 days after delivery, 18F-FDG-PET was performed to evaluate therapeutic efficacy. ResultsWith intravenous delivery, Ad-pSurvivin-TSTA-TRAIL-FL showed no measurable liver tumor FL signal on day 1 after delivery, but showed better therapeutic efficacy than Ad-pCMV-FL on day 7 (PET tumor/liver ratio, 3.5 ± 0.58 vs 6.0 ± 0.71; P = .02). With intratumoral delivery, Ad-pSurvivin-TSTA-TRAIL-FL showed positive FL signal from all tumors and better therapeutic efficacy than Ad-pCMV-FL on day 7 (2.4 ± 0.50 vs 5.4 ± 0.78; P = .01). In addition, intratumoral delivery of Ad-pSurvivin-TSTA-TRAIL-FL demonstrated significant decrease in tumoral viability compared with intravenous delivery (2.4 ± 0.50 vs 3.5 ± 0.58; P = .03). ConclusionsIntratumoral delivery of a transcriptionally targeted therapeutic vector for amplifying tumor-specific effect demonstrated better transduction efficiency and therapeutic efficacy for liver cancer than systemic delivery, and may lead to improved therapeutic outcome for future clinical practice.

CD8+ T cell response mediates the therapeutic effects of oncolytic adenovirus in an immunocompetent mouse model

The role of anti-tumor immune responses in oncolytic adenoviral therapy has not been well studied due to lack of efficacious tumor model in immunocompetent mice. Here, we evaluated the contributions of immune components to the therapeutic effects of oncolytic adenoviruse in an immunocompetent murine tumor model permissive for infection and replication of adenovirus. We found that CD8+ T cells were critical mediator for antitumor efficacy by oncolytic adenovirus. Intratumoral viral therapy induced intensive infiltration of CD8+ T cells in tumor, increased tumor-specific IFN-γ (interferon-γ) production and CTL (cytotoxic T lymphocyte) activity of lymphocytes, and generated a long-term tumor-specific immune memory. Boosting CD8+ T cell responses by agonistic anti-4-1BB (cluster differentiation 137, CD137) antibody showed synergistic anticancer effects with oncolytic virotherapy. Our results provide insight into antitumor mechanisms of oncolytic adenovirus in addition to their direct oncolytic effect.

Fermented Mistletoe Extract as a Multimodal Antitumoral Agent in Gliomas

In Europe, commercially available extracts from the white-berry mistletoe (Viscum album L.) are widely used as a complementary cancer therapy. Mistletoe lectins have been identified as main active components and exhibit cytotoxic effects as well as immunomodulatory activity. Since it is still not elucidated in detail how mistle toe extracts such as ISCADOR communicate their effects, we analyzed the mechanisms that might be responsible for their antitumoral function on a molecular and functional level. ISCADOR-treated glioblastoma (GBM) cells down-regulate central genes involved in glioblastoma progression and malignancy such as the cytokine TGF-β and matrix-metalloproteinases. Using in vitro glioblastoma/immune cell co-cultivation assays as well as measurement of cell migration and invasion, we could demonstrate that in glioblastoma cells, lectin-rich ISCADOR M and ISCADOR Q significantly enforce NK-cell-mediated GBM cell lysis. Beside its immune stimulatory effect, ISCADOR reduces the migratory and invasive potential of glioblastoma cells. In a syngeneic as well as in a xenograft glioblastoma mouse model, both pretreatment of tumor cells and intratumoral therapy of subcutaneously growing glioblastoma cells with ISCADOR Q showed delayed tumor growth. In conclusion, ISCADOR Q, showing multiple positive effects in the treatment of glioblastoma, may be a candidate for concomitant treatment of this cancer.

Calreticulin promotes tumor lymphocyte infiltration and enhances the antitumor effects of immunotherapy by up‐regulating the endothelial expression of adhesion molecules

Tumor-induced angiogenesis has been shown to suppress immune responses. One mechanism is to suppress leukocyte-endothelial cell interaction by down-regulating the expression of adhesion molecules, such as intercellular adhesion molecule (ICAM)-1, vascular cell adhesion molecule (VCAM)-1 and E-selectin on the tumor endothelium, which enables tumor cells to escape immune surveillance. Calreticulin (CRT), a chaperone protein mainly located in the endoplasmic reticulum, has been shown to exert anti-angiogenic activity and inhibit tumor growth. Here, we demonstrate that in addition to inhibiting angiogenesis, CRT also enhances the expression of both ICAM-1 and VCAM-1 on tumor endothelial cells. This expression results in enhanced leukocyte-endothelial cell interactions and increased lymphocyte infiltration into tumors. Therefore, combining intramuscular CRT gene transfer with intratumoral cytokine gene therapies significantly improves the antitumor effects of immunotherapy by markedly increasing the levels of tumor-infiltrating lymphocytes. This combined treatment increased the levels of infiltrating lymphocytes to those achieved using four times the cytokine dosage. The combined therapy also resulted in lower levels of immunosuppressive molecules and higher levels of activated T-cells in the tumor microenvironment than immunotherapy alone. In conclusion, this study describes a new antitumor mechanism of CRT that involves the up-regulation of tumor endothelial adhesion molecules and the enhanced infiltration of tumor-specific lymphocytes. Thus, CRT treatment can make tumor cells more vulnerable to immunotherapy and improve the therapeutic efficacy of immunotherapy.

Percutaneous Intratumoral Injection With Pingyangmycin Lipiodol Emulsion for the Treatment of Recurrent Sacrococcygeal Chordomas

This study describes fluoroscopy-guided percutaneous intratumoral injection therapy (PIIT) with a pingyangmycin lipiodol emulsion in the management of recurrent sacrococcygeal chordomas after surgical excision. Seven patients underwent a total of 22 treatment sessions (3–4 sessions per patient); treatment responses were evaluated clinically, and lesion size was determined using computed tomography (CT). Over 10–26 months of follow-up, tumor sizes and visual analogue scale (VAS) scores of all patients were decreased. No patients had complications during the follow-up period. Preliminary results showed that PIIT with pingyangmycin lipiodol emulsion under fluoroscopic guidance is effective and safe and may be considered as a treatment option.

SU‐E‐I‐121: MRI Detection of Dominant Intraprostatic Lesions and Correlation with Tumor Microenvironment

Purpose: To determine if MRI is suitable for planning of intratumoral radionuclide therapy of prostatic cancer by correlation of MRI imaging parameters to histologic parameters Methods: All statistical analysis was performed using GraphPad Prism version 5.00 for Windows, GraphPad Software, San Diego California USA, www.graphpad.com. A total of 22 tumors from 10 patients who underwent 3T endorectal MRI and subsequent prostatectomy were investigated retrospectively. Differences in raw T2 signal, T2 signal ratios, apparent diffusion coefficient (ADC), and ADC ratios were tested for significance using one‐way ANOVA for normal tissue, index lesions and secondary lesions as a function of specific pathology parameters (percent stroma, percent glandular area, percent cellularity, glandular area, glandular perimeter, stromal width). Bonferroni multiple comparisons test was performed to test differences in index and secondary tumors as a part of ANOVA analysis. Pearsons correlation was performed for each MRI parameter and pathology parameter based on tissue type as well. Results: Normal tissue MRI parameters were found to be significantly different (p <.05) from index and secondary tumors for all pathologic parameters. Index and secondary lesions showed no significant differences from one another (p<.05). Significant correlations were seen for normal tissue, index and secondary lesions for T2 signal intensity ratios and raw T2 when looking at glandular area and percent luminal space (p <.05, r >.66). Other moderate to strong correlations were seen for ADC and secondary lesions (r < −.58) Conclusions: These results show that MRI has he ability to distinguish between index lesions and secondary lesions as well as informing on the tissue microenvironment. A larger, randomized prospective trial should be performed to further test this hypothesis. MRI will play a central role in successfully planning intratumoral radionuclide therapy of prostatic index lesions in the future.

Abstract 2669: Radiosensitizing effect of intratumoral mIL-12 electrogene therapy in murine sarcoma

Abstract Due to the limited success rate of conventional therapeutic approaches to soft tissue sarcomas, new treatment modalities, such as gene therapy, are being evaluated as adjuvant therapies to conventional methods. Electrogene therapy with interleukin-12 (IL-12) has been proved very effective due to the induction of adaptive immunity and innate resistance as well as anti-angiogenic action. The aim of our study was to evaluate the radiosensitizing effect of single and repetitive intratumoral (i.t.) mIL-12 electrogene therapy (mIL-12 EGT) in murine sarcoma. Solid SA-1 fibrosarcoma tumors were induced in the back of A/J mice by subcutaneous injection of tumor cells. The first therapeutic approach consisted of a single i.t. mIL-12 EGT (20 µg of plasmid DNA), followed by a single dose (10 Gy) or graded doses (2.5 – 70 Gy) of local tumor irradiation 24 h later. In the second therapeutic approach, tumor irradiation was followed by two additional i.t. mIL-12 EGT, on the third and fifth day. The tumor growth delay, complete response rates and local tumor control assay (TCD50 assay) were used as a measure of antitumor effectiveness of the therapies and skin damage in the irradiation field was also scored. Tumor and serum concentrations of cytokines mIL-12 and mouse interferon γ (mIFNγ) were measured using ELISA. Therapies with i.t. mIL-12 EGT demonstrated good antitumor effects in murine SA-1 sarcoma tumors and also a radiosensitizing effect when combined with irradiation. Both antitumor and radiosensitizing effects were more pronounced when using repetitive rather than single i.t. mIL-12 EGT. Single i.t. mIL-12 EGT resulted in 7% complete responses and, when combined with single dose irradiation, it produced a radiosensitizing effect resulting in 21% complete responses. In both cases we also detected significantly elevated levels of cytokines mIL-12 and mIFNγ in tumors but no significant increase of both cytokine levels in the serum. Repetitive i.t. mIL-12 EGT alone resulted in 75% complete responses and, when combined with single-dose irradiation, produced the most pronounced potentiation of the radiation response resulting in 100% complete responses of the treated tumors. The TCD50 value was 30 Gy for irradiation alone, and 13.8 Gy for i.t. mIL-12 EGT combined with irradiation. Therefore, the dose-modifying factor of a single i.t. mIL-12 EGT in murine SA-1 sarcoma was 2.17. At the same level of skin damage, dry desquamation, in combined therapy a 44% higher probability of local tumor control than with irradiation alone was observed. The results of our study demonstrate that i.t. mIL-12 EGT is an efficient treatment modality for treatment of soft tissue sarcoma alone and as adjuvant therapy to single dose irradiation. This is due to its pronounced radiosensitizing effect caused by boosting the immune response of the organism. The dose-modifying factor of a single i.t. mIL-12 EGT in murine SA-1 sarcoma was 2.17. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2669. doi:10.1158/1538-7445.AM2011-2669

Abstract 3650: Anti-MIF gene therapy of murine mammary tumor results in decreased tumor metastasis and reduction of MDSC (Myeloid derived suppressor cells)

Abstract Introduction: Macrophage migration inhibitory factor (MIF) is a multifunctional cytokine which has been shown to be secreted by a variety of tumors and results in autocirne tumor growth. The purpose of this study is to determine the effect of down-regulation of MIF expression on primary tumor growth, metastasis and cellular mediators of immune suppression. Methods: Ten days post 4T1-luciferase tumor cells injection into Balb/c mice, nanoparticles with anti-MIF siRNA and controls (PBS and nanoparticles with scrambled siRNA) were directly injected into established tumor daily for 7 days. At different time points, tumor sizes were measured, tumor metastases in live mice were imaged, tumors and blood samples were harvested. The gene expression in tumor slides and 4T1 cells from tumor digest were investigated by FACS and immunofluorescence microscopy. The total number of circulating MDSCs and MDSCs expressing suppressive cytokines were determined by FACS. Results: We have developed a non-viral gene carrier system that contains a biodegradable β-glucan (10 Kda) backbone and protonable imidazole at the reducing end. The nanoparticles at N/P of 8 lead to the high uptake efficiency by 4T1 cells. The imidazole has buffering capacity to induce endosomal escape. Once released into cytosol, the biodegradable glucan shell of nanoparticles is slowly oxidized and degraded so that the siRNAs are sustained released and the target gene MIF is sustained reduced. Applying this intratumoral anti-MIF gene therapy in vivo, we have made the following observations: 1) MIF protein is highly expressed in 4T1 mammary tumors and can be efficiently blocked in vivo; 2) Reduction of MIF production in established tumor in vivo results in reduced primary tumor size and metastasis; 3) Reduction of MIF production in 4T1 tumor cells in vitro results in reduced MIF secretion in 4T1-conditioned medium and enhanced tumor cell apoptosis and death; 4) Reduction of MIF production in established tumor results in a significant reduction in the total number of circulating MDSCs by 37.6%. The MDSCs expressing immune suppressive cytokines including TGF-β, IL-10 and IL-1β was reduced by 43%, 47% and 70%. Significance: Although MIF appears to have recognized autocrine growth activity in a variety of tumors, development of therapeutics directed at MIF have been problematic because it serves a number of functions in normal biological processes. Targeted reduction of MIF by intratumoral injection of siRNA using a non-viral nanoparticle technology alleviates the difficulty of gene delivery. In addition, since the down-regulation of tumor-secreted MIF may reduce mammary tumor-induced immune suppression by reducing MDSCs, the local therapy can result in a favorable systemic antitumor effect by enhancing host immunity. This study represents a first step in developing this therapeutic approach. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3650. doi:10.1158/1538-7445.AM2011-3650

716 EUS Guided Intra-Tumoral Gemcitabine Therapy for Locally Advanced and Metastatic Pancreatic Cancer

One of the biggest hurdles to progress in chemoradiotherapy of pancreatic cancer (PC) is the lack of penetration of therapeutic agents into the cancer due to tumor desmoplasia. Despite great optimism for new regimens, outcomes have not meaningfully changed and new therapies are needed to improve local control. We hypothesized that direct intratumoral injection of chemotherapy would enhance the direct drug specific effects as well as boost local radiation effect within the tumor bed. In patients with biopsy proven PC and performance status of 0-1 we sought to determine the toxicity and efficacy of EUS guided intratumoral fine needle injection (FNI) of gemcitabine when administered as one time induction therapy given prior to conventional multimodality therapy. 36 patients with locally advanced or metastatic PC underwent EUS FNI with gemcitabine (40mg/ml) after being deemed unresectable (n=34), a poor operative candidate (n=1), or after patient refusal of resection (n=1). At 4-14 days following EUS FNI, and prior to initiating standard chemoradiotherapy, a helical CT, hematology labs, hepatic and renal chemistries, and clinical follow-up were obtained to assess initial toxicity. The primary endpoint was toxicity. Secondary endpoints included time to progression, ability to downstage, and survival (6 and 12 month). We enrolled 36 PC patients (mean age 65; range 42-86 years; 19M/17F), 33 of whom had stage III/IV disease. A mean of 2.7 needle passes (range 1-4) and mean total volume of 2.5 ml (range 0.7-7.0) was injected per patient, corresponding to an intratumoral injection of 90 mg (range 28-280 mg) of gemcitabine. There were no EUS FNI procedure-related complications or grade 3 or higher adverse events associated with intratumoral therapy. Currently 16 patients are deceased with a mean survival of 6.4 mo (range 2.8-14.6). The remaining 20 patients are alive at a mean of 11.8 mo (range 4.8-18.2) following treatment initiation. Survival at 6 months and 1 year is 76% and 46% for 34 and 28 evaluable patients, respectively. Three patients initially deemed unresectable have been downstaged and undergone R0 resection. They include patients staged T3N1M0, T4N0M0, and T3N1M1 who underwent pancreatoduodenectomy, distal pancreatectomy, and pancreatoduodenectomy, respectively. In these patients, there is no evidence of disease recurrence at 10.2, 14.6, and 6.0mo, respectively. Our study suggests the feasibility and safety of intratumoral EUS FNI of gemcitabine for pancreatic cancer. These data offer promise in terms of tumor downstaging and impact on survival. Continued follow-up of current patients and additional study are needed to verify these findings and to address other key radiographic and clinical endpoints, and to refine the technique and clarify the potential role relative to standard therapy.Tabled 1StagePtsExpected Overall Survival (mo) Without Surgery based on National Cancer DatabaseStudy Patients-Deceased (n=16); Time and Death; Mean (Range)Study Patients-Alive (n=20), Duration in months: Mean (Range)IA06.8n=0n=0IB16.1n=05.2 (n=1)IIA16.2n=016.6 (n=1)IIB16.79.8 (n=1)n=0III227.26.7 (3.4-14.6) (n=12)12.2 (n=10)IV112.55.0 (n=3)10.7 (n=8) Open table in a new tab

Abstract No. 190: A phase I/II safety and efficacy study of intratumoral light-activated drug therapy using talaporfin sodium in patients with inoperable hepatocellular carcinoma

Talaporfin sodium (TS) is a light-activated small molecule in development for the treatment of solid tumors. TS was evaluated in combination with a proprietary drug activator in a phase I/II safety and efficacy trial in patients with hepatocellular carcinoma (HCC). To our knowledge, this is the first clinical study of light-activated drug therapy in patients with HCC.

Cancer Electrogene Therapy with Interleukin-12

Electrogene therapy combines administration of plasmid DNA into tissue followed by local application of electric pulses. In electrogene therapy with interleukin-12 (IL-12), different routes of administration, different doses of plasmid DNA and different protocols for delivery of electric pulses were evaluated in numerous preclinical studies. Antitumor effectiveness was tested in different types of primary tumors, distantly growing tumors and induced metastases. Intratumoral IL-12 electrogene therapy has been proved to be very effective in local tumor control, having also a systemic effect. Intramuscular and peritumoral IL-12 electrogene therapy had also a pronounced systemic effect and when combined with other treatment strategies resulted in tumor cures. Antitumor effectiveness of IL-12 electrogene therapy is due to the induction of adaptive immunity and innate resistance and anti-angiogenic action. Translation of preclinical studies into clinical trials in human and veterinary oncology has started with encouraging results that would hopefully lead to further investigation of this therapy, also in combination with other cancer treatment modalities.

Intratumoral Therapy of Glioblastoma Multiforme Using Genetically Engineered Transferrin for Drug Delivery

Glioblastoma multiforme (GBM) is the most common and lethal primary brain tumor with median survival of only 12 to 15 months under the current standard of care. To both increase tumor specificity and decrease nonspecific side effects, recent experimental strategies in the treatment of GBM have focused on targeting cell surface receptors, including the transferrin (Tf) receptor, that are overexpressed in many cancers. A major limitation of Tf-based therapeutics is the short association of Tf within the cell to deliver its payload. We previously developed two mutant Tf molecules, K206E/R632A Tf and K206E/K534A Tf, in which iron is locked into each of the two homologous lobes. Relative to wild-type Tf, we showed enhanced delivery of diphtheria toxin (DT) from these mutants to a monolayer culture of HeLa cells. Here, we extend the application of our Tf mutants to the treatment of GBM. In vitro treatment of Tf mutants to a monolayer culture of glioma cells showed enhanced cellular association as well as enhanced delivery of conjugated DT. Treatment of GBM xenografts with mutant Tf-conjugated DT resulted in pronounced regression in vivo, indicating their potential use as drug carriers.

Abstract 3824: Induction of potent antitumor immunity by intratumoral injection of mT-bet transduced dendritic cells

Abstract T-bet is a T-box transcription factor (Tbx21) previously identified as a master regulator of Type-1 polarization in T cells. We have previously demonstrated that ectopic expression of hT-bet converts dendritic cells (DC) into Type 1-biasing APCs. In the current study, we investigated whether mouse DC overexpressing mT-bet promote superior Type 1 anti-tumor T cell responses when applied as an intratumoral therapy. CD11c+ DCs were then purified and transduced with recombinant adenovirus to express high levels of mT-bet. DC. mT-bet stimulated stronger Tc1 and Th1 T cell responses when compared with control DC and retained the capacity to kill and then uptake tumor cells/debris, allowing for the cross-priming of specific T cell responses in vitro. Surprisingly, in contrast to human DC. T-bet, CD40L-activated DC. mT-bet produced elevated levels (vs. control DC) of IL-12p70/TNF-α. DC. mT-bet exerted potent anti-tumor activity in the (day 7 established) s.c. CMS4 sarcoma after intratumoral administration. These anti-tumor effects were mediated primarily by Tc1 cells, however, both CD4+ T cells and NK cells appeared to also be involved in therapeutic response. In situ imaging suggested that DC. mT-bet treatment inhibited the accumulation of CD11b+Gr1+ myeloid-derived suppressor cells and tumor-associated vascular structures. Collectively, these findings suggest that DC. mT-bet-based therapy has the potential to promote superior Type-1 anti-tumor immunity in vivo. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3824.

Abstract 2411: Intratumoral dendritic cell vaccine as neo-adjuvant immunotherapy of high-risk soft tissue sarcoma patients undergoing external beam radiation therapy (EBRT): Prospective phase I/II clinical trial

Abstract Cell death inducing external beam radiation (EBRT) combined with experimental intratumoral injection of dendritic cells (DC) showed promise by initiating anti-autologous tumor-cell immune responses in mice. Here we performed first clinical trial testing this hypothesis in patients with large high-grade soft tissue sarcomas (STS). This type of cancer has a significant (>50%) risk of progressing to distant metastases. Patients with clinical stage T2N0M0 or T3N0M0 high-grade STS of the extremity/trunk/chest wall were treated with standard neo-adjuvant EBRT 5040 cGy / 180 cGy coordinated with experimental DC therapy consisting of DC progenitor apheresis, ex-vivo expansion and culture, and 4 × intratumoral injections of 10 million DC. Autologous DC product was performed according to standardized GMP laboratory procedures. T cell function was assessed by EliSpot (measuring IFN-γ production) and proliferation (thymidine uptake) in addition to phenotyping of peripheral blood mononuclear cells by flow cytometry. Clinically, targeted accrual was reached with eighteen patients completing neo-adjuvant EBRT with experimental intratumoral DC therapy. All patients have completed full immunologic assessment. Ten patients (56%) were induced to produce significant immune responses against autologous tumor cell lysates or/and survivin antigens as determined using ELISPOT assays or cell proliferation. Interestingly, some of these responses persisted even 30 weeks after start of treatment. Extensive post treatment T cell infiltration was detected within tumors. No clinical toxicity has been observed. Suggestive clinical outcome on 14 patients followed 1 year reveal progress in 1/7 patients with robust response (detected at more than one time point), and 4/7 patients with weak (detected at one time point at the beginning of the study) or no response respectively. Thus, this data is promising for planning future combined treatment approaches as radiation therapy/immunotherapy. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2411.

Intratumoral cancer chemotherapy and immunotherapy: opportunities for nonsystemic preoperative drug delivery.

The recent literature documents the growing interest in local intratumoral chemotherapy as well as systemic preoperative chemotherapy with evidence for improved outcomes using these therapeutic modalities. Nevertheless, with few exceptions, the conventional wisdom and standard of care for clinical and surgical oncology remains surgery followed by radiation and/or systemic chemotherapy, as deemed appropriate based on clinical findings. This, in spite of the fact that the toxicity of conventional systemic chemotherapy and immunotherapy affords limited effectiveness and frequently compromises the quality of life for patients. Indeed, with systemic chemotherapy, the oncologist (and the patient) often walks a fine line between attempting tumour remission with prolonged survival and damaging the patient's vital functions to the point of death. In this context, it has probably been obvious for more than 100 years, due in part to the pioneering work of Ehrlich (1878), that targeted or localized drug delivery should be a major goal of chemotherapy. However, there is still only limited clinical use of nonsystemic intratumoral chemotherapy for even those high mortality cancers which are characterized by well defined primary lesions i.e. breast, colorectal, lung, prostate, and skin. There has been a proliferation of intratumoral chemotherapy and immunotherapy research during the past two to three years. It is therefore the objective of this review to focus much more attention upon intratumoral therapeutic concepts which could limit adverse systemic events and which might combine clinically feasible methods for localized preoperative chemotherapy and/or immunotherapy with surgery. Since our review of intratumoral chemoimmunotherapy almost 20 years ago (McLaughlin & Goldberg 1983), there have been few comprehensive reviews of this field; only one of broad scope (Brincker 1993), three devoted specifically to gliomas (Tomita 1991; Walter et al. 1995; Haroun & Brem 2000), one on hepatomas (Venook 2000), one concerning veterinary applications (Theon 1998), and one older review of dermatological applications (Goette 1981). However, none have shed light on practical opportunities for combining intratumoral therapy with subsequent surgical resection. Given the state-of-the-art in clinical and surgical oncology, and the advances that have been made in intratumoral drug delivery, minimally invasive tumour access i.e. fine needle biopsy, new drugs and drug delivery systems, and preoperative chemotherapy, it is timely to present a review of studies which may suggest future opportunities for safer, more effective, and clinically practical non-systemic therapy.

A Case of Hepatocellular Carcinoma with Pulmonary Metastases Treated Successfully with a Combination of Repeated Hepatic Arterial Infusion Epirubicin and Cisplatin Chemotherapy and Systemic Low-Dose Infusion of 5-Fluorouracil

We report a case of hepatocellular carcinoma (HCC) with pulmonary metastases treated with repeated hepatic arterial infusion chemotherapy (HAIC) comprising epirubicin and cisplatin, and systemic infusion of 5-fluorouracil (a modified EC/F protocol), which led to complete remission. A 49-year-old man with compensated liver cirrhosis experienced intrahepatic recurrence of HCC with extensive lung metastases. The modified EC/F therapeutic protocol, which was applied at the tenth cycle every 4-5 weeks, resulted in disappearance of the pulmonary metastases and normalization of serum α-fetoprotein levels. A single small HCC lesion was found in the left lobe of the liver 13 months after the final chemotherapy session. HAIC with the same regimen was conducted again, followed by percutaneous intratumoral chemoinjection therapy with 5-fluorouracil and interferon-γ. Thereafter, there was no evidence of recurrence in either the liver or the lung, as evidenced by image analysis and expression of tumor markers. The disease-free intervals for the liver and lung were 41 and 54 months, respectively.

Local and systemic antitumor effect of intratumoral and peritumoral IL-12 electrogene therapy on murine sarcoma

Soft tissue sarcomas pose a challenge for successful treatment with conventional therapeutic methods, therefore newer therapeutic approaches are considered. In this study, we evaluated the antitumor effect of IL-12 electrogene therapy (EGT) on murine SA-1 fibrosarcoma. The Therapeutic plasmid was injected either intratumorally into subcutaneous SA-1 nodules or intradermally into the peritumoral region. We achieved a remarkable local and systemic antitumor effect with both approaches after single plasmid DNA application, with significant intratumoral and systemic production of IL-12 and IFN-γ. Intratumoral IL-12 EGT resulted in over 90% complete response rate of the treated tumors with 60% of cured mice being resistant to challenge with SA-1 tumor cells. Peritumoral EGT resulted in a lower complete response rate (16%), with significant growth delay of remaining tumors. Both therapies also resulted in significant inhibition of growth of untreated tumors, growing simultaneously at a distant site. These data suggest that IL-12 EGT may be useful in the treatment of soft tissue sarcomas, exerting a local and systemic antitumor effect.

Interventional Endoscopic Ultrasound: The Next Frontier in Gastrointestinal Endoscopy

Endoscopic ultrasound (EUS) has become well established as a diagnostic modality in gastrointestinal cancer staging. It offers high-resolution imaging and fine-needle biopsy, which is essential in tumor and nodal staging of gastrointestinal cancers. In the recent decade, however, many therapeutic applications of EUS have become possible. Currently, interventional EUS endoscopy involves celiac plexus neurolysis, pseudocyst drainage, and intratumoral fine-needle injection therapy for inoperable pancreatic malignancy. Emerging techniques include the accurate endoscopic delivery of radioactive beads to localize tumor therapy as well as other therapies, such as radiofrequency ablation or cryotherapy. Diagnostic and therapeutic access to the biliary tree and pancreatic duct is increasingly being used successfully in failed endoscopic retrograde cholangiopancreatography (ERCP) procedures. This review discusses these procedures and several evolving future applications, including vascular access and EUS-guided enteral anastomosis.