Akseli Hemminki

Publications (119)706.11 Total impact

• Article: Oncolytic Adenovirus With Temozolomide Induces Autophagy and Antitumor Immune Responses in Cancer Patients.

  Ilkka Liikanen, Laura Ahtiainen, Mari Lm Hirvinen, Simona Bramante, Vincenzo Cerullo, Petri Nokisalmi, Otto Hemminki, Iulia Diaconu, Sari Pesonen, Anniina Koski, Lotta Kangasniemi, Saila K Pesonen, Minna Oksanen, Leena Laasonen, Kaarina Partanen, Timo Joensuu, Fang Zhao, Anna Kanerva, Akseli Hemminki
  [show abstract] [hide abstract]
  ABSTRACT: Oncolytic adenoviruses and certain chemotherapeutics can induce autophagy and immunogenic cancer cell death. We hypothesized that the combination of oncolytic adenovirus with low-dose temozolomide (TMZ) is safe, effective, and capable of inducing antitumor immune responses. Metronomic low-dose cyclophosphamide (CP) was added to selectively reduce regulatory T-cells. Preclinically, combination therapy inhibited tumor growth, increased autophagy, and triggered immunogenic cell death as indicated by elevated calreticulin, adenosine triphosphate (ATP) release, and nuclear protein high-mobility group box-1 (HMGB1) secretion. A total of 41 combination treatments given to 17 chemotherapy-refractory cancer patients were well tolerated. We observed anti- and proinflammatory cytokine release, evidence of virus replication, and induction of neutralizing antibodies. Tumor cells showed increased autophagy post-treatment. Release of HMGB1 into serum-a possible indicator of immune response-increased in 60% of treatments, and seemed to correlate with tumor-specific T-cell responses, observed in 10/15 cases overall (P = 0.0833). Evidence of antitumor efficacy was seen in 67% of evaluable treatments with a trend for increased survival over matched controls treated with virus only. In summary, the combination of oncolytic adenovirus with low-dose TMZ and metronomic CP increased tumor cell autophagy, elicited antitumor immune responses, and showed promising safety and efficacy.Molecular Therapy (2013); doi:10.1038/mt.2013.51.
  Molecular Therapy 04/2013; · 6.87 Impact Factor
• Article: Anti-viral and anti-tumor T-cell immunity in patients treated with GMCSF coding oncolytic adenovirus.

  Anna Kanerva, Petri Nokisalmi, Iulia Diaconu, Anniina Koski, Vincenzo Cerullo, Ilkka Liikanen, Siri Tähtinen, Minna Oksanen, Raita Heiskanen, Saila K Pesonen, Tima Joensuu, Tuomo Alanko, Kaarina P L Partanen, Leena Laasonen, Kalevi Kairemo, Sari Pesonen, Lotta Kangasniemi, Akseli Hemminki
  [show abstract] [hide abstract]
  ABSTRACT: Purpose and study design: Multiple injections of oncolytic adenovirus could enhance immunological response. In the first part of this manuscript the focus was on immunological aspects. 60 patients previously naive to oncolytic virus, and who had white blood cells available, were treated. 39/60 were assessed after a single virus administration, while 21/60 received a "serial treatment" consisting of 3 injections within 10 weeks. In the second part, we focused on 115 patients treated with a GMCSF coding capsid chimeric adenovirus, CGTG-102. RESULTS: Following serial treatment, both increase and decrease in anti-tumor T-cells in blood were seen more frequently, findings which are compatible with induction of T-cell immunity and trafficking of T-cells to tumors, respectively. Safety was good in both groups. In 115 patients treated with CGTG-102 (Ad5/3-D24-GMCSF), median overall survival was 111 days following single and 277 days after serial treatment in non-randomized comparison. Switching the virus capsid for avoiding neutralizing antibodies, in a serial treatment featuring three different viruses, did not impact safety or efficacy. A correlation between anti-viral and anti-tumor T-cells was seen (P = 0.001), suggesting that viral oncolysis can result in epitope spreading and breaking of tumor-associated immunological tolerance. Alternatively, some patients may be more susceptible to induction of T-cell immunity and/or trafficking. CONCLUSIONS: These results provide the first human data linking anti-viral immunity with anti-tumor immunity, implying that oncolytic viruses could have an important role in cancer immunotherapy.
  Clinical Cancer Research 03/2013; · 7.74 Impact Factor
• Article: 2-weekly versus 3-weekly docetaxel to treat castration-resistant advanced prostate cancer: a randomised, phase 3 trial.

  Pirkko-Liisa Kellokumpu-Lehtinen, Ulrika Harmenberg, Timo Joensuu, Ray McDermott, Petteri Hervonen, Claes Ginman, Marjaana Luukkaa, Paul Nyandoto, Akseli Hemminki, Sten Nilsson, John McCaffrey, Raija Asola, Taina Turpeenniemi-Hujanen, Fredrik Laestadius, Tiina Tasmuth, Katinka Sandberg, Maccon Keane, Ilari Lehtinen, Tiina Luukkaala, Heikki Joensuu
  [show abstract] [hide abstract]
  ABSTRACT: BACKGROUND: Docetaxel administered every 3 weeks is a standard treatment for castration-resistant advanced prostate cancer. We hypothesised that 2-weekly administration of docetaxel would be better tolerated than 3-weekly docetaxel in patients with castration-resistant advanced prostate cancer, and did a prospective, multicentre, randomised, phase 3 study to compare efficacy and safety. METHODS: Eligible patients had advanced prostate cancer (metastasis, a prostate-specific-antigen test result of more than 10·0 ng/mL, and WHO performance status score of 0-2), had received no chemotherapy (except with estramustine), had undergone surgical or chemical castration, and had been referred to a treatment centre in Finland, Ireland, or Sweden. Enrolment and treatment were done between March 1, 2004, and May 31, 2009. Randomisation was done centrally and stratified by centre and WHO performance status score of 0-1 vs 2. Patients were assigned 75 mg/m(2) docetaxel intravenously on day 1 of a 3-week cycle, or 50 mg/m(2) docetaxel intravenously on days 1 and 15 of a 4-week cycle. 10 mg oral prednisolone was administered daily to all patients. The primary endpoint was time to treatment failure (TTTF). We assessed data in the per-protocol population. This study is registered with ClinicalTrials.gov, number NCT00255606. FINDINGS: 177 patients were randomly assigned to the 2-weekly docetaxel group and 184 to the 3-weekly group. 170 patients in the 2-weekly group and 176 in the 3-weekly group were included in the analysis. The 2-weekly administration was associated with significantly longer TTTF than was 3-weekly administration (5·6 months, 95% CI 5·0-6·2 vs 4·9 months, 4·5-5·4; hazard ratio 1·3, 95% CI 1·1-1·6, p=0·014). Grade 3-4 adverse events occurred more frequently in the 3-weekly than in the 2-weekly administration group, including neutropenia (93 [53%] vs 61 [36%]), leucopenia (51 [29%] vs 22 [13%]), and febrile neutropenia (25 [14%] vs six [4%]). Neutropenic infections were reported more frequently in patients who received docetaxel every 3 weeks (43 [24%] vs 11 [6%], p=0·002). INTERPRETATION: Administration of docetaxel every 2 weeks seems to be well tolerated in patients with castration-resistant advanced prostate cancer and could be a useful option when 3-weekly single-dose administration is unlikely to be tolerated. FUNDING: Sanofi.
  The lancet oncology 01/2013; · 14.47 Impact Factor
• Article: MicroRNA-Mediated Suppression of Oncolytic Adenovirus Replication in Human Liver.

  Erkko Ylösmäki, Sergio Lavilla-Alonso, Sari Jäämaa, Markus Vähä-Koskela, Taija Af Hällström, Akseli Hemminki, Johanna Arola, Heikki Mäkisalo, Kalle Saksela
  [show abstract] [hide abstract]
  ABSTRACT: MicroRNAs (miRNAs) are important and ubiquitous regulators of gene expression that can suppress their target genes by translational inhibition as well as mRNA destruction. Cell type-specific miRNA expression patterns have been successfully exploited for targeting the expression of experimental and therapeutic gene constructs, for example to reduce pathogenic effects of cancer virotherapy in normal tissues. In order to avoid liver damage associated with systemic or intrahepatic delivery of oncolytic adenoviruses we have introduced the concept of suppressing adenovirus replication in hepatic cells by inserting target elements for the liver-specific miR122 into the viral genome. Here we show using ex vivo cultured tissue specimens that six perfectly complementary miR122 target sites in the 3' untranslated region of the viral E1A gene are sufficient in the absence of any other genetic modifications to prevent productive replication of serotype 5 adenovirus (Ad5) in normal human liver. This modification did not compromise the replicative capacity of the modified virus in cancer tissue derived from a colon carcinoma liver metastasis or its oncolytic potency in a human lung cancer xenograft mouse model. Unlike wild-type Ad5, the modified virus did not result in increased serum levels of liver enzymes in infected mice. These results provide a strong preclinical proof of concept for the use of miR122 target sites for reducing the risk of liver damage caused by oncolytic adenoviruses, and suggest that ectopic miR122 target elements should be considered as an additional safety measure included in any therapeutic virus or viral vector posing potential hazard to the liver.
  PLoS ONE 01/2013; 8(1):e54506. · 4.09 Impact Factor
• Article: Mutation of the fiber shaft heparan sulphate binding site of a 5/3 chimeric adenovirus reduces liver tropism.

  Anniina Koski, Eerika Karli, Anja Kipar, Sophie Escutenaire, Anna Kanerva, Akseli Hemminki
  [show abstract] [hide abstract]
  ABSTRACT: Natural tropism to the liver is a major obstacle in systemic delivery of adenoviruses in cancer gene therapy. Adenovirus binding to soluble coagulation factors and to cellular heparan sulphate proteoglycans via the fiber shaft KKTK domain are suggested to cause liver tropism. Serotype 5 adenovirus constructs with mutated KKTK regions exhibit liver detargeting, but they also transduce tumors less efficiently, possibly due to altered fiber conformation. We constructed Ad5/3lucS*, a 5/3 chimeric adenovirus with a mutated KKTK region. The fiber knob swap was hypothesized to facilitate tumor transduction. This construct was studied with or without additional coagulation factor ablation. Ad5/3lucS* exhibited significantly reduced transduction of human hepatic cells in vitro and mouse livers in vivo. Combination of coagulation factor ablation by warfarinization to Ad5/3lucS* seemed to further enhance liver detargeting. Cancer cell transduction by Ad5/3lucS* was retained in vitro. In vivo, viral particle accumulation in M4A4-LM3 xenograft tumors was comparable to controls, but Ad5/3lucS* transgene expression was nearly abolished. Coagulation factor ablation did not affect tumor transduction. These studies set the stage for further investigations into the effects of the KKTK mutation and coagulation factor ablation in the context of 5/3 serotype chimerism. Of note, the putative disconnect between tumor transduction and transgene expression could prove useful in further understanding of adenovirus biology.
  PLoS ONE 01/2013; 8(4):e60032. · 4.09 Impact Factor
• Article: Oncolytic adenoviruses: A potent form of tumor immunovirotherapy.

  Vincenzo Cerullo, Markus Vähä-Koskela, Akseli Hemminki
  [show abstract] [hide abstract]
  ABSTRACT: The therapeutic efficacy of oncolytic viruses including adenovirus has been thought to depend mostly on direct viral destruction of tumor cells. However, this view has changed with the discovery that oncolysis can also induce innate and antigen-specific adaptive immunity against the tumor. Here we summarize our findings from cancer patients.
  Oncoimmunology. 09/2012; 1(6):979-981.
• Article: Ad3-hTERT-E1A, a fully serotype 3 oncolytic adenovirus, in patients with chemotherapy refractory cancer.

  Otto Hemminki, Iulia Diaconu, Vincenzo Cerullo, Saila K Pesonen, Anna Kanerva, Timo Joensuu, Kalevi Kairemo, Leena Laasonen, Kaarina Partanen, Lotta Kangasniemi, Andre Lieber, Sari Pesonen, Akseli Hemminki
  [show abstract] [hide abstract]
  ABSTRACT: Twenty-five patients with chemotherapy refractory cancer were treated with a fully serotype 3-based oncolytic adenovirus Ad3-hTERT-E1A. In mice, Ad3 induced higher amounts of cytokines but less liver damage than Ad5 or Ad5/3. In humans, the only grade 3 adverse reactions were self-limiting cytopenias and generally the safety profile resembled Ad5-based oncolytic viruses. Patients that had been previously treated with Ad5 viruses presented longer lasting lymphocytopenia but no median increase in Ad3-specific T-cells in blood, suggesting immunological activity against antigens other than Ad3 hexon. Frequent alterations in antitumor T-cells in blood were seen regardless of previous virus exposure. Neutralizing antibodies against Ad3 increased in all patients, whereas Ad5 neutralizing antibodies remained stable. Treatment with Ad3-hTERT-E1A resulted in re-emergence of Ad5 viruses from previous treatments into blood and vice versa. Signs of possible efficacy were seen in 11/15 (73%) patients evaluable for tumor markers, four of which were treated only intravenously. Particularly promising results were seen in breast cancer patients and especially those receiving concomitant trastuzumab. Taken together, Ad3-hTERT-E1A seems safe for further clinical testing or development of armed versions. It offers an immunologically attractive alternative, with possible pharmacodynamic differences and a different receptor compared to Ad5.
  Molecular Therapy 08/2012; 20(9):1821-30. · 6.87 Impact Factor
• Article: An Oncolytic Adenovirus Enhanced for Toll-like Receptor 9 Stimulation Increases Antitumor Immune Responses and Tumor Clearance.

  Vincenzo Cerullo, Iulia Diaconu, Valentina Romano, Mari Hirvinen, Matteo Ugolini, Sophie Escutenaire, Sirkka-Liisa Holm, Anja Kipar, Anna Kanerva, Akseli Hemminki
  [show abstract] [hide abstract]
  ABSTRACT: Oncolytic viruses represent a multifaceted tool for cancer treatment. In addition to specific killing of cancer cells (oncolysis), these agents also provide danger signals prompting the immune system to stimulate an antitumor immune response. To increase adenovirus adjuvancy, we engineered the genome of Ad5D24 by inserting 18 immunostimulatory islands (Ad5D24-CpG). The toxicity and immunogenicity profile of Ad5D24-CpG showed that the safety of the maternal virus was retained. The efficacy of the CpG-enriched virus was assessed in a xenograft model of lung cancer where a significant increase in antitumor effect was seen in comparison with controls. When the experiment was repeated in animal depleted of natural killer (NK) cells, Ad5D24-CpG lost its advantage. The same was seen when Toll-like receptor (TLR)9 was blocked systemically. In a syngeneic model of melanoma (B16-OVA), we observed a significant increase of OVA-specific T cells and a decrease of activation of myeloid-derived suppressor cells in Ad5D24-CpG-treated mice. In conclusion, we have generated the first genetically modified oncolytic adenovirus backbone able to enhance TLR9-stimulation for increased antitumor activity.
  Molecular Therapy 07/2012; · 6.87 Impact Factor
• Article: A new human DSG2-transgenic mouse model for studying the tropism and pathology of human adenoviruses.

  Hongjie Wang, Ines Beyer, Jonas Persson, Hui Song, ZongYi Li, Maximilian Richter, Hua Cao, Ruan van Rensburg, Xiaoying Yao, Kelly Hudkins, Roma Yumul, Xiao-Bing Zhang, Mujun Yu, Pascal Fender, Akseli Hemminki, André Lieber
  [show abstract] [hide abstract]
  ABSTRACT: We have recently reported that a group of human adenoviruses (HAdVs) uses desmoglein 2 (DSG2) as a receptor for infection. Among these are the widely distributed serotypes HAdV-B3 and HAdV-B7, as well as a newly emerged strain derived from HAdV-B14. These serotypes do not infect rodent cells and could not up until now be studied in small-animal models. We therefore generated transgenic mice containing the human DSG2 locus. These mice expressed human DSG2 (hDSG2) at a level and in a pattern similar to those found for humans and nonhuman primates. As an initial application of hDSG2-transgenic mice, we used a green fluorescent protein (GFP)-expressing HAdV-B3 vector (Ad3-GFP) and studied GFP transgene expression by quantitative reverse transcription-PCR (qRT-PCR) and immunohistochemistry subsequent to intranasal and intravenous virus application. After intranasal application, we found efficient transduction of bronchial and alveolar epithelial cells in hDSG2-transgenic mice. Intravenous Ad3-GFP injection into hDSG2-transgenic mice resulted in hDSG2-dependent transduction of epithelial cells in the intestinal and colon mucosa. Our findings give an explanation for clinical symptoms associated with infection by DSG2-interacting HAdVs and provide a rationale for using Ad3-derived vectors in gene therapy.
  Journal of Virology 03/2012; 86(11):6286-302. · 5.40 Impact Factor
• Article: Immune response is an important aspect of the antitumor effect produced by a CD40L-encoding oncolytic adenovirus.

  Iulia Diaconu, Vincenzo Cerullo, Mari L M Hirvinen, Sophie Escutenaire, Matteo Ugolini, Saila K Pesonen, Simona Bramante, Suvi Parviainen, Anna Kanerva, Angelica S I Loskog, Aristides G Eliopoulos, Sari Pesonen, Akseli Hemminki
  [show abstract] [hide abstract]
  ABSTRACT: Oncolytic adenovirus is an attractive platform for immunotherapy because virus replication is highly immunogenic and not subject to tolerance. Although oncolysis releases tumor epitopes and provides costimulatory danger signals, arming the virus with immunostimulatory molecules can further improve efficacy. CD40 ligand (CD40L, CD154) induces apoptosis of tumor cells and triggers several immune mechanisms, including a T-helper type 1 (T(H)1) response, which leads to activation of cytotoxic T cells and reduction of immunosuppression. In this study, we constructed a novel oncolytic adenovirus, Ad5/3-hTERT-E1A-hCD40L, which features a chimeric Ad5/3 capsid for enhanced tumor transduction, a human telomerase reverse transcriptase (hTERT) promoter for tumor selectivity, and human CD40L for increased efficacy. Ad5/3-hTERT-E1A-hCD40L significantly inhibited tumor growth in vivo via oncolytic and apoptotic effects, and (Ad5/3-hTERT-E1A-hCD40L)-mediated oncolysis resulted in enhanced calreticulin exposure and HMGB1 and ATP release, which were suggestive of immunogenicity. In two syngeneic mouse models, murine CD40L induced recruitment and activation of antigen-presenting cells, leading to increased interleukin-12 production in splenocytes. This effect was associated with induction of the T(H)1 cytokines IFN-γ, RANTES, and TNF-α. Tumors treated with Ad5/3-CMV-mCD40L also displayed an enhanced presence of macrophages and cytotoxic CD8(+) T cells but not B cells. Together, our findings show that adenoviruses coding for CD40L mediate multiple antitumor effects including oncolysis, apoptosis, induction of T-cell responses, and upregulation of T(H)1 cytokines.
  Cancer Research 03/2012; 72(9):2327-38. · 7.86 Impact Factor
• Article: Oncolytic immunotherapy of advanced solid tumors with a CD40L-expressing replicating adenovirus: assessment of safety and immunologic responses in patients.

  Sari Pesonen, Iulia Diaconu, Lotta Kangasniemi, Tuuli Ranki, Anna Kanerva, Saila K Pesonen, Ulrike Gerdemann, Ann M Leen, Kalevi Kairemo, Minna Oksanen, Elina Haavisto, Sirkka-Liisa Holm, Aila Karioja-Kallio, Satu Kauppinen, Kaarina P L Partanen, Leena Laasonen, Tima Joensuu, Tuomo Alanko, Vincenzo Cerullo, Akseli Hemminki
  [show abstract] [hide abstract]
  ABSTRACT: The immunosuppressive environment of advanced tumors is a primary obstacle to the efficacy of immunostimulatory and vaccine approaches. Here, we report an approach to arm an oncolytic virus with CD40 ligand (CD40L) to stimulate beneficial immunologic responses in patients. A double-targeted chimeric adenovirus controlled by the hTERT promoter and expressing CD40L (CGTG-401) was constructed and nine patients with progressing advanced solid tumors refractory to standard therapies were treated intratumorally. No serious adverse events resulting in patient hospitalization occurred. Moderate or no increases in neutralizing antibodies were seen, suggesting effective Th1 immunologic effects. An assessment of the blood levels of virus indicated 17.5% of the samples (n = 40) were positive at a low level early after treatment, but not thereafter. In contrast, high levels of virus, CD40L, and RANTES were documented locally at the tumor. Peripheral blood mononuclear cells were analyzed by IFN-γ ELISPOT analysis and induction of both survivin-specific and adenovirus-specific T cells was seen. Antitumor T-cell responses were even more pronounced when assessed by intracellular cytokine staining after stimulation with tumor type-specific peptide pools. Of the evaluable patients, 83% displayed disease control at 3 months and in both cases in which treatment was continued the effect was sustained for at least 8 months. Injected and noninjected lesions responded identically. Together, these findings support further clinical evaluation of CGTG-401.
  Cancer Research 02/2012; 72(7):1621-31. · 7.86 Impact Factor
• Article: A phase II trial of gefitinib in patients with rising PSA following radical prostatectomy or radiotherapy.

  Greetta Joensuu, Timo Joensuu, Nina Nupponen, Mirja Ruutu, Juhani Collan, Sari Pesonen, Akseli Hemminki
  Acta oncologica (Stockholm, Sweden) 01/2012; 51(1):130-3. · 2.27 Impact Factor
• Source

  Available from: Anu J Airaksinen

  Article: SPECT/CT imaging of hNIS-expression after intravenous delivery of an oncolytic adenovirus and 131I.

  Maria Rajecki, Mirkka Sarparanta, Tanja Hakkarainen, Mikko Tenhunen, Iulia Diaconu, Venla Kuhmonen, Kalevi Kairemo, Anna Kanerva, Anu J Airaksinen, Akseli Hemminki
  [show abstract] [hide abstract]
  ABSTRACT: Oncolytic adenoviruses can be engineered for better tumor selectivity, gene delivery and be armed for imaging and concentrating radionuclides into tumors for synergistic oncolysis. We constructed Ad5/3-hTERT-hNIS where replication is controlled by hTERT-promoter. Ad5/3-hTERT-hNIS expresses hNIS for imaging of transgene expression and for treatment of infected tumors by radioiodine. Ad5/3-hTERT-hNIS efficiently killed prostate cancer cells and induced iodine uptake in vitro and in vivo after intratumoral virus administration. Survival of mice treated with intravenous Ad5/3-hTERT-hNIS significantly prolonged survival over mock or radioiodine only but the combination of virus with radioiodine was not more effective than virus alone. Temporal and spatial changes in hNIS-expression during therapy were detected with SPECT, demonstrating feasibility of evaluation of the combination therapy with hNIS-expressing adenoviruses and radioiodide.
  PLoS ONE 01/2012; 7(3):e32871. · 4.09 Impact Factor
• Article: Oncolytic adenoviruses for cancer immunotherapy: data from mice, hamsters, and humans.

  Vincenzo Cerullo, Anniina Koski, Markus Vähä-Koskela, Akseli Hemminki
  [show abstract] [hide abstract]
  ABSTRACT: Adenovirus is one of the most commonly used vectors for gene therapy and two products have already been approved for treatment of cancer in China (Gendicine(R) and Oncorine(R)). An intriguing aspect of oncolytic adenoviruses is that by their very nature they potently stimulate multiple arms of the immune system. Thus, combined tumor killing via oncolysis and inherent immunostimulatory properties in fact make these viruses in situ tumor vaccines. When further engineered to express cytokines, chemokines, tumor-associated antigens, or other immunomodulatory elements, they have been shown in various preclinical models to induce antigen-specific effector and memory responses, resulting both in full therapeutic cures and even induction of life-long tumor immunity. Here, we review the state of the art of oncolytic adenovirus, in the context of their capability to stimulate innate and adaptive arms of the immune system and finally how we can modify these viruses to direct the immune response toward cancer.
  Advances in Cancer Research 01/2012; 115:265-318. · 4.46 Impact Factor
• Article: Verapamil results in increased blood levels of oncolytic adenovirus in treatment of patients with advanced cancer.

  Anniina Koski, Mari Raki, Petri Nokisalmi, Ilkka Liikanen, Lotta Kangasniemi, Timo Joensuu, Anna Kanerva, Sari Pesonen, Ramon Alemany, Akseli Hemminki
  [show abstract] [hide abstract]
  ABSTRACT: Calcium channel blockers including verapamil have been proposed to enhance release and antitumor efficacy of oncolytic adenoviruses in preclinical studies but this has not been studied in humans before. Here, we studied if verapamil leads to increased replication of oncolytic adenovirus in cancer patients, as measured by release of virions from tumor cells into the systemic circulation. The study was conducted as a matched case-control study of advanced cancer patients treated with oncolytic adenoviruses with or without verapamil. We observed that verapamil increased mean virus titers present in blood after treatment (P < 0.05). The frequency or severity of adverse events was not increased, nor were cytokine responses or neutralizing antibody levels different between groups. Signs of possible treatment-related clinical benefits were observed in both groups, but there was no significant difference in responses or survival. Thus, our data suggests that the combination of verapamil with oncolytic adenoviruses is safe and well tolerated. Moreover, verapamil treatment seems to result in higher virus titers in blood, indicating enhanced overall replication in tumors. A randomized trial is needed to confirm these findings and to study if enhanced replication results in benefits to patients.
  Molecular Therapy 11/2011; 20(1):221-9. · 6.87 Impact Factor
• Article: Radiation-induced upregulation of gene expression from adenoviral vectors mediated by DNA damage repair and regulation.

  Petri Nokisalmi, Maria Rajecki, Sari Pesonen, Sophie Escutenaire, Rabah Soliymani, Mikko Tenhunen, Laura Ahtiainen, Akseli Hemminki
  [show abstract] [hide abstract]
  ABSTRACT: In the present study, we evaluated the combination of replication-deficient adenoviruses and radiotherapy in vitro. The purpose of the present study was to analyze the mechanism of radiation-mediated upregulation of adenoviral transgene expression. Adenoviral transgene expression (luciferase or green fluorescent protein) was studied with and without radiation in three cell lines: breast cancer M4A4-LM3, prostate cancer PC-3MM2, and lung cancer LNM35/enhanced green fluorescent protein. The effect of the radiation dose, modification of the viral capsid, and five different transgene promoters were studied. The cellular responses were studied using mass spectrometry and immunofluorescence analysis. Double strand break repair was modulated by inhibitors of heat shock protein 90, topoisomerase-I, and DNA protein kinase, and transgene expression was measured. We found that a wide range of radiation doses increased adenoviral transgene expression regardless of the cell line, transgene, promoter, or viral capsid modification. Treatment with adenovirus, radiation, and double strand break repair inhibitors resulted in persistence of double strand breaks and subsequent increases in adenovirus transgene expression. Radiation-induced enhancement of adenoviral transgene expression is linked to DNA damage recognition and repair. Radiation induces a global cellular response that results in increased production of RNA and proteins, including adenoviral transgene products. This study provides a mechanistic rationale for combining radiation with adenoviral gene delivery.
  International journal of radiation oncology, biology, physics 10/2011; 83(1):376-84. · 4.59 Impact Factor
• Article: Effects of capsid-modified oncolytic adenoviruses and their combinations with gemcitabine or silica gel on pancreatic cancer.

  Lotta Kangasniemi, Suvi Parviainen, Tommi Pisto, Mika Koskinen, Mika Jokinen, Tuula Kiviluoto, Vincenzo Cerullo, Harry Jalonen, Anniina Koski, Anna Kangasniemi, Anna Kanerva, Sari Pesonen, Akseli Hemminki
  [show abstract] [hide abstract]
  ABSTRACT: Conventional cancer treatments often have little impact on the course of advanced pancreatic cancer. Although cancer gene therapy with adenoviruses is a promising developmental approach, the primary receptor is poorly expressed in pancreatic cancers which might compromise efficacy and thus targeting to other receptors could be beneficial. Extended stealth delivery, combination with standard chemotherapy or circumvention of host antiadenoviral immune response might improve efficacy further. In this work, capsid-modified adenoviruses were studied for transduction of cell lines and clinical normal and tumor tissue samples. The respective oncolytic viruses were tested for oncolytic activity in vitro and in vivo. Survival was studied in a peritoneally disseminated pancreas cancer model, with or without concurrent gemcitabine while silica implants were utilized for extended intraperitoneal virus delivery. Immunocompetent mice and Syrian hamsters were used to study the effect of silica mediated delivery on antiviral immune responses and subsequent in vivo gene delivery. Capsid modifications selectively enhanced gene transfer to malignant pancreatic cancer cell lines and clinical samples. The respective oncolytic viruses resulted in increased cell killing in vitro, which translated into a survival benefit in mice. Early proinfammatory cytokine responses and formation of antiviral neutralizing antibodies was partially avoided with silica implants. The implant also shielded the virus from pre-existing neutralizing antibodies, while increasing the pancreas/liver gene delivery ratio six-fold. In conclusion, capsid modified adenoviruses would be useful for testing in pancreatic cancer trials. Silica implants might increase the safety and efficacy of the approach.
  International Journal of Cancer 08/2011; 131(1):253-63. · 5.44 Impact Factor
• Article: Induction of interferon pathways mediates in vivo resistance to oncolytic adenovirus.

  Ilkka Liikanen, Vladia Monsurrò, Laura Ahtiainen, Mari Raki, Tanja Hakkarainen, Iulia Diaconu, Sophie Escutenaire, Otto Hemminki, João D Dias, Vincenzo Cerullo, Anna Kanerva, Sari Pesonen, Daniela Marzioni, Marco Colombatti, Akseli Hemminki
  [show abstract] [hide abstract]
  ABSTRACT: Oncolytic adenoviruses are an emerging experimental approach for treatment of tumors refractory to available modalities. Although preclinical results have been promising, and clinical safety has been excellent, it is also apparent that tumors can become virus resistant. The resistance mechanisms acquired by advanced tumors against conventional therapies are increasingly well understood, which has allowed development of countermeasures. To study this in the context of oncolytic adenovirus, we developed two in vivo models of acquired resistance, where initially sensitive tumors eventually gain resistance and relapse. These models were used to investigate the phenomenon on RNA and protein levels using two types of analysis of microarray data, quantitative reverse transcriptase-polymerase chain reaction and immunohistochemistry. Interferon (IFN) signaling pathways were found upregulated and Myxovirus resistance protein A (MxA) expression was identified as a marker correlating with resistance, while transplantation experiments suggested a role for tumor stroma in maintaining resistance. Furthermore, pathway analysis suggested potential therapeutic targets in oncolytic adenovirus-resistant cells. Improved understanding of the antiviral phenotype causing tumor recurrence is of key importance in order to improve treatment of advanced tumors with oncolytic adenoviruses. Given the similarities between mechanisms of action, this finding might be relevant for other oncolytic viruses as well.
  Molecular Therapy 07/2011; 19(10):1858-66. · 6.87 Impact Factor
• Article: Multimerization of adenovirus serotype 3 fiber knob domains is required for efficient binding of virus to desmoglein 2 and subsequent opening of epithelial junctions.

  Hongjie Wang, ZongYi Li, Roma Yumul, Stephanie Lara, Akseli Hemminki, Pascal Fender, André Lieber
  [show abstract] [hide abstract]
  ABSTRACT: Recently, we identified desmoglein 2 (DSG2) as the main receptor for a group of species B adenoviruses (Ads), including Ad3, a serotype that is widely distributed in the human population (H. Wang et al., Nat. Med. 17:96-104, 2011). In this study, we have attempted to delineate structural details of the Ad3 interaction with DSG2. For CAR- and CD46-interacting Ad serotypes, attachment to cells can be completely blocked by an excess of recombinant fiber knob protein, while soluble Ad3 fiber knob only inefficiently blocks Ad3 infection. We found that the DSG2-interacting domain(s) within Ad3 is formed by several fiber knob domains that have to be in the spatial constellation that is present in viral particles. Based on this finding, we generated a small recombinant, self-dimerizing protein containing the Ad3 fiber knob (Ad3-K/S/Kn). Ad3-K/S/Kn bound to DSG2 with high affinity and blocked Ad3 infection. We demonstrated by confocal immunofluorescence and transmission electron microscopy analyses that Ad3-K/S/Kn, through its binding to DSG2, triggered the transient opening of intercellular junctions in epithelial cells. The pretreatment of epithelial cells with Ad3-K/S/Kn resulted in increased access to receptors that are localized in or masked by epithelial junctions, e.g., CAR or Her2/neu. Ad3-K/S/Kn treatment released CAR from tight junctions and thus increased the transduction of epithelial cells by a serotype Ad5-based vector. Furthermore, the pretreatment of Her2/neu-positive breast cancer cells with Ad3-K/S/Kn increased the killing of cancer cells by the Her2/neu-targeting monoclonal antibody trastuzumab (Herceptin). This study widens our understanding of how Ads achieve high avidity to their receptors and the infection of epithelial tissue. The small recombinant protein Ad3-K/S/Kn has practical implications for the therapy of epithelial cancer and gene/drug delivery to normal epithelial tissues.
  Journal of Virology 07/2011; 85(13):6390-402. · 5.40 Impact Factor
• Source

  Available from: Thomas Rudge

  Article: A new generation of serotype chimeric infectivity-enhanced conditionally replicative adenovirals: the safety profile of ad5/3-Δ24 in advance of a phase I clinical trial in ovarian cancer patients.

  Kenneth H Kim, Michael J Ryan, James E Estep, Brock M Miniard, Thomas L Rudge, James O Peggins, Trevor L Broadt, Minghui Wang, Meredith A Preuss, Gene P Siegal, Akseli Hemminki, Raymond D Harris, Rosemarie Aurigemma, David T Curiel, Ronald D Alvarez
  [show abstract] [hide abstract]
  ABSTRACT: Conditionally replicative adenoviral (CRAd) virotherapy represents a promising therapeutic approach for cancer. We have demonstrated that a serotype chimeric adenoviral 5/3 fiber-knob modification achieves enhanced ovarian cancer infectivity, conditional replication, and oncolytic activity. This study evaluated the safety of intraperitoneal (IP) Ad5/3-Δ24 in advance of a phase I clinical trial in gynecologic cancers. Syrian hamster cohorts were treated with IP Ad5/3-Δ24 or control buffer for 3 consecutive days and euthanized on study days 8, 17, 57, and 89. Blood and tissue samples were harvested from each animal. For biodistribution studies, presence and quantitation of viral levels within samples were determined via quantitative polymerase chain reaction. For safety studies, animals were assessed for adverse vector-related tissue or laboratory effects. In the biodistribution study, low levels of Ad5/3-Δ24 DNA were noted outside of the abdominal cavity. Viral DNA levels in tissues obtained from the peritoneal cavity peaked at day 8 and declined thereafter. In the safety study, no specific histopathologic changes were attributable to virus administration. Hematologic findings noted in the 1 × 10(11) viral particles (vp)/dose group on Days 4 and/or 8 were indicative of an Ad5/3-Δ24-specific generalized inflammatory response; these findings resolved by day 56. The no observable adverse effect level was determined to be 1 × 10(10) vp/dose. This study elucidates the safety profile of IP administration of the serotype chimeric infectivity-enhanced CRAd, Ad5/3-Δ24, and provides guidance for a planned phase I trial for patients with recurrent gynecologic cancers.
  Human gene therapy 07/2011; 22(7):821-8. · 4.20 Impact Factor