Search

Oncolytic viruses (OVs) have been shown to induce anti-cancer immunity and enhance cancer immunotherapies, such as immune checkpoint inhibitor therapies. OV therapies can be further improved by arming OVs with immunostimulatory molecules, including various cytokines or chemokines. Here, we have developed a novel adenovirus encoding two immunostimulatory molecules: cluster of differentiation 40 ligand (CD40L) and tumor necrosis factor receptor superfamily member 4 ligand (OX40L). This novel virus, designated VALO-D102, is designed to activate both innate and adaptive immune responses against tumors. CD40L affects the innate side by licensing antigen-presenting cells to drive CD8+ T cell responses, and OX40L increases clonal expansion and survival of CD8+ T cells and formation of a larger pool of memory T cells. VALO-D102 and its murine surrogate VALO-mD901, expressing murine OX40L and CD40L, were used in our previously developed PeptiCRAd cancer vaccine platform. Intratumoral administration of PeptiCRAd significantly increased tumor-specific T cell responses, reduced tumor growth, and induced systemic anti-cancer immunity in two mouse models of melanoma. In addition, PeptiCRAd therapy, in combination with anti-PD-1 immune checkpoint inhibitor therapy, significantly improved tumor growth control as compared to either monotherapy alone., Graphical Abstract, Ylösmäki et al. develop an oncolytic adenovirus expressing immunostimulatory cytokines OX40L and CD40L. This virus was tested in a peptide-based cancer vaccine platform called PeptiCRAd. Intratumoral administration of PeptiCRAd induced systemic and intratumoral tumor-specific T cell responses, reduced tumor growth, and sensitized tumors to immune checkpoint inhibitor therapy.

Docetaxel-based chemotherapy is effective in metastatic gastric and gastro-oesophageal junction adenocarcinoma. This study reports on the safety and efficacy of the docetaxel-based triplet FLOT (fluorouracil plus leucovorin, oxaliplatin and docetaxel) as a perioperative therapy for patients with locally advanced, resectable tumours.

Neoepitope-specific T-cell responses have been shown to induce durable clinical responses in patients with advanced cancers. We explored the recognition patterns of tumor-infiltrating T lymphocytes (TILs) from patients with glioblastoma multiforme (GBM), the most fatal form of tumors of the central nervous system. Whole-genome sequencing was used for generating DNA sequences representing the entire spectrum of ‘private’ somatic mutations in GBM tumors from five patients, followed by 15-mer peptide prediction and subsequent peptide synthesis. For each mutated peptide sequence, the wildtype sequence was also synthesized and individually co-cultured with autologous GBM TILs, which had been expanded in vitro with a combination of interleukin (IL)-2, IL-15 and IL-21. After seven days of culture, interferon gamma (IFN-γ), tumor necrosis factor alpha (TNF-α) and/or IL-17A production was measured by ELISA in culture supernatants, and used as an epitope-specific immune response readout. Mutated peptides that induced a strong cytokine response were considered to contain legitimate neoepitopes. TILs from 5/5 patients with GBM exhibited specific immune reactivity profiles to the nominal target peptides, defined by IFN-γ and/or TNF-α production, as well as IL-17A. Neoepitopes, defined by mutated peptides inducing IFN-γ and/or TNF-α production without or only minimal reactivity to the wildtype sequences, were found for each individual patient. CD8+ TILs dominated the patients’ responses to private neoepitopes. The present study shows that neoepitope-specific TIL reactivity constitutes an important arm of anti-tumor immune responses in patients with GBM, and thus a powerful tool for developing next-generation personalized immunotherapies.

SPIRIT 2013 Checklist. (PDF 167 kb)

All Items from the World Health Trial Registration Data Set. (PDF 74 kb)

Late stage cancer is often associated with reduced immune recognition and a highly immunosuppressive tumor microenvironment. The presence of tumor infiltrating lymphocytes (TILs) and specific gene-signatures prior to treatment are linked to good prognosis, while the opposite is true for extensive immunosuppression. The use of adenoviruses as cancer vaccines is a form of active immunotherapy to initialise a tumor-specific immune response that targets the patient’s unique tumor antigen repertoire. We report a case of a 68-year-old male with asbestos-related malignant pleural mesothelioma who was treated in a Phase I study with a granulocyte-macrophage colony‑stimulating factor (GM-CSF)-expressing oncolytic adenovirus, Ad5/3-D24-GMCSF (ONCOS-102). The treatment resulted in prominent infiltration of CD8C lymphocytes to tumor, marked induction of systemic antitumor CD8C T-cells and induction of Th1- type polarization in the tumor. These results indicate that ONCOS-102 treatment sensitizes tumors to other immunotherapies by inducing a T-cell positive phenotype to an initially T-cell negative tumor.

Jährlich erkranken etwa 425 000 Menschen in Deutschland an Krebs. Die Tendenz ist steigend: Experten gehen davon aus, dass die Zahl der Neuerkrankungen bis zum Jahr 2030 um 50 Prozent zunehmen wird. Doch zu dieser schlechten Nachricht gibt es auch eine gute: Körperliche Aktivität und Sport können das allgemeine Risiko, an bestimmten Krebsformen zu erkranken, vermindern. Dazu zählen vor allem Darmkrebs sowie der nach den Wechseljahren auftretende Brust- und Gebärmutterschleimhautkrebs. Aber auch wer schon erkrankt ist, kann sein Wohlbefinden und Selbstvertrauen durch spezielle Bewegungsprogramme, wie sie an der Goethe-Universität entwickelt werden, steigern. Selbst die Leiden von Patienten mit fortgeschrittenen Krebserkrankungen lassen sich auf diese Weise lindern. Denn Bewegung beeinflusst nicht nur die unmittelbar tumorbedingten Symptome, sondern auch therapiebedingte Nebenwirkungen, insbesondere die der Chemotherapie.

Antibody-based cancer immunotherapy relies on the identification and characterization of target antigens and the development of potent antibodies recognizing the target. Here we report the expression analysis and molecular characterization of the differentiation antigen NY-BR-1, which we previously identified by using the SEREX (serological analysis of recombinant cDNA expression libraries) method. Corroborating methodologies, including mRNA quantitation and immunoblotting show that NY-BR-1 is strongly expressed in >70% of 129 breast tumors. Application of a NY-BR-1 specific antibody demonstrated NY-BR-1 expression in primary and metastastic breast cancers. In contrast, most of the breast cancer cell lines tested, expressed only low NY-BR-1 levels. Importantly, confocal microscopy revealed that ectopically expressed NY-BR-1 localizes to the cytoplasm and the cell membrane. NY-BR-1 localization in breast cancer specimens was also confirmed by immunohistochemistry. Bioinformatic analysis and deletion mutagenesis further show that NY-BR-1 membrane localization is mediated by 2 cis-active membrane targeting domains. Biochemical surface labeling and FACS analysis of live cells further characterize NY-BR-1 as a transmembrane protein, which can be specifically recognized by the anti-NY-BR-1 antibody on the surface of vital cells. The strong expression of NY-BR-1 in breast tumors, its cytoplasmic and membrane localization and accessibility to an ectopically applied antibody now suggest to pursue NY-BR-1 as a potential target for antibody-based therapies in breast cancer patients.

The treatment results of many cancers have improved, but there are still challenges, such as metastatic cancers that mostly lack curative treatments. Oncolytic adenoviruses provide a new therapeutic option. Adenoviruses can be modified to replicate specifically in cancer cells and thereby causing the death of cancer cells (oncolysis). Adenovirus treatments are well tolerated and they can be combined with conventional treatments. In the pre-clinical section of this thesis we evaluated the combination of adenoviruses and radiotherapy. We studied the mechanism of radiation-mediated upregulation of adenoviral transgenes. Three cancer cell lines were used: M4A4-LM3 (breast cancer), PC-3MM2 (prostate cancer) and LNM35/eGFP (lung cancer). Cancer cells were exposed to radiation and infected with various replication-deficient adenoviruses. The transgene expression with or without radiation was evaluated. The radiation response was also modulated by DNA protein kinase-, heat shock protein 90- and topoisomerase-I inhibitor. Radiation increased adenovirus transgene expression (luciferase or GFP) regardless of the transgene, promoter, cancer cell line or radiation dose. We showed that enhancement of transgene expression is mediated through genotoxic stress regulation and repair. This synergy can be used for therapeutic benefit. In the clinical section 157 cancer patients with advanced solid tumors were treated with different oncolytic adenoviruses. The safety, efficacy and immunological responses of treatments were evaluated. The safety was monitored by clinical chemistry, cytokine measurements and CTCAE analysis for adverse events. Immunological responses were quantified by neutralizing antibodies, viral genomes in the blood and measuring responses in tumor- and adenovirus-specific T-cells. RECIST analysis and tumor markers were applied to treatment efficacy analysis. We also compared the safety and efficacy of a serial treatment scheme, three rounds of virus, to a single treatment. Adenovirus treatments were well tolerated and commonly detected clinical adverse events were of grade 2 or less including injection or tumor site pain, nausea, fever and fatigue. Six out of 157 treated patients (3.8 %) experienced a grade 4 adverse event. Serious adverse events were seen in 11 patients (7.0 %). No treatment related deaths occurred. According to RECIST analysis disease control (= stable disease or better) was seen in 40.0 - 74.0 % of patients and a stabilization or a decrease in tumor markers for 23.1 - 70.6 % of patients. Serial treatment was as well tolerated as single treatment, but results suggested a better median survival even though statistical significance was not reached. These findings represent a good justification for forthcoming clinical trials. Useiden syöpien hoitotulokset ovat parantuneet viime vuosina, mutta kuitenkaan useimpiin levinneisiin syöpiin ei ole tarjolla parantavia hoitoja. Uusia hoitomenetelmiä tulee siis jatkuvasti kehittää. Onkolyyttiset adenovirukset ovat eräs lupaava uusi hoitomuoto. Adenovirukset ovat yleisiä luonnossa esiintyviä viruksia, jotka aiheuttavat ihmisissä lieviä sairauksia kuten hengitystieinfektioita. Onkolyyttisiä adenoviruksia on muokattu niin, että ne tuhoavat vain syöpäsoluja (onkolyysi) säästäen terveitä soluja. Hoidot tällaisilla viruksilla ovat yleensä potilaille hyvin siedettyjä ja niitä voidaan yhdistää myös muihin hoitoihin. Tässä väitöskirjassa tutkittiin sädehoidon ja adenovirusten yhteisvaikutuksia rinta, eturauhas- ja keuhkosyöpäsoluissa. Adenovirukset kykenevät kuljettamaan mukanaan syöpäsoluihin geenejä joiden tuottamilla proteiineilla on syöpäsoluja tuhoava vaikutus. Tällaisia geenejä kutsutaan siirtogeeneiksi. Tutkimuksessa arvioitiin miten sädehoito vaikuttaa näiden adenovirusten siirtogeenien määrään. Siirtogeenien ja niiden tuottamien proteiinien määrä oli suurempi sellaisissa soluissa joita oli säteilytetty ennen adenovirusten antamista. Näiden tulosten perusteella vaikuttaa olevan hyödyllistä yhdistää adenovirusgeeniterapia sekä sädehoito. Tulokset luovat biologisen perustan yhdistelmähoidolle, joka voi parhaimmillaan tulevaisuudessa parantaa hoidettujen potilaiden ennustetta. Väitöskirjan kliinisessä osassa tutkittiin adenovirushoidon turvallisuutta, tehoa ja immunologista vastetta levinnyttä syöpää sairastavilla potilailla (157 potilasta). Turvallisuutta arvioitiin verikokein ja hoitoihin liittyvät haittavaikutukset luokiteltiin vakavuuden mukaan tasoihin 1 5. Hoidon immunologista vastetta potilaissa arvioitiin mm. mittaamalla viruksen ja T-valkosolujen määrää verenkierrossa. Hoidon tehoa arvioitiin radiologisella kuvantamisella sekä syöpämerkkiaineilla. Lisäksi sarjahoitoa, joka tarkoittaa kolmea peräkkäistä virushoitoa, verrattiin kertahoitoon. Adenovirushoidot olivat hyvin siedettyjä ja yleisesti haitat olivat lieviä tasoa 1 - 2 kuten kiputuntemukset, pahoinvointi, kuume ja väsymys. Kuudella potilaalla (3.8 %) havaittiin tasoa 4 oleva haittavaikutus ja vakavia haittoja 11 potilaalla (7.0 %). Hoitoon liittyviä kuolemia ei ollut. Radiologinen hoitovaste saavutettiin 40.0 - 74.0 %:lle potilaista ja kasvainmerkkiaineilla mitattuna 23.1 - 70.6 %:lle. Sarjahoito oli yhtä turvallinen kuin kertahoito, mutta sillä oli suotuisa vaikutus potilaiden elossaoloaikaan, joskaan ero ei ollut tilastollisesti merkittävä. Tuloksemme ovat lupaavia ja siksi Pohjois-Euroopan ensimmäinen kliininen tutkimus onkolyyttisellä viruksella on aloitettu.