Drug development in oncology assisted by noninvasive optical imaging.

International audienceEarly and accurate detection of tumors, like the development of targeted treatments, is a major field of research in oncology. The generation of specific vectors, capable of transporting a drug or a contrast agent to the primary tumor site as well as to the remote (micro-) metastasis would be an asset for early diagnosis and cancer therapy. Our goal was to develop new treatments based on the use of tumor-targeted delivery of large biomolecules (DNA, siRNA, peptides, or nanoparticles), able to induce apoptosis while dodging the specific mechanisms developed by tumor cells to resist this programmed cell death. Nonetheless, the insufficient effectiveness of the vectorization systems is still a crucial issue. In this context, we generated new targeting vectors for drug and biomolecules delivery and developed several optical imaging systems for the follow-up and evaluation of these vectorization systems in live mice. Based on our recent work, we present a brief overview of how noninvasive optical imaging in small animals can accelerate the development of targeted therapeutics in oncology

AGuIX® from bench to bedside-Transfer of an ultrasmall theranostic gadolinium-based nanoparticle to clinical medicine

International audienceAGuIX® are sub-5 nm nanoparticles made of a polysiloxane matrix and gadolinium chelates. This nanoparticle has been recently accepted in clinical trials in association with radiotherapy. This review will summarize the principal preclinical results that have led to first in man administration. No evidence of toxicity has been observed during regulatory toxicity tests on two animal species (rodents and monkeys). Biodistributions on different animal models have shown passive uptake in tumours due to enhanced permeability and retention effect combined with renal elimination of the nanoparticles after intravenous administration. High radiosensitizing effect has been observed with different types of irradiations in vitro and in vivo on a large number of cancer types (brain, lung, melanoma, head and neck…). The review concludes with the second generation of AGuIX nanoparticles and the first preliminary results on human

Vectorisation de biomolécules pour l'imagerie et la thérapie des cancers

Early and accurate detection of tumors, as well as the development of targeted-treatments are major field of research in oncology. The generation of specific vectors, capable of addressing a drug or a contrast agent to the primary tumor site as well as in the remote (micro-) metastasis would be thus a major asset for early diagnosis and cancer therapy. In this double goal of diagnosis and therapy, nanoparticles emerged as promising candidates for the establishment of therapeutic protocols, including targeting, diagnosis with optical imaging techniques and cancer therapy. In this context, our laboratory developed 2D and 3D optical imaging devices, designed for small animal imaging, to enable the long term non-invasive follow-up of the in vivo biodistribution of fluorescent probes and nanoparticles. The characterization of the behavior of different nanoparticles (lipidic nanocapsules, gadolinium oxide nanoparticles and gold nanoparticles) with various surface functionalizations was carried out in vitro and in vivo on different tumor models. The results show the influence of the functionalization and the size of nanoparticles on their in vivo biodistribution and on their way of elimination. On the other hand, the functionalization of the nanoparticles by the RGD motif allows a specific targeting of cells expressing the alpha v beta 3 integrin in vitro and improves the in vivo accumulation in tumors.Le diagnostic précis et précoce des tumeurs, ainsi que le développement de thérapies ciblées sont deux axes principaux de recherche en cancérologie. La génération de vecteurs spécifiques capables d'adresser un agent de contraste et/ou un médicament au niveau de la tumeur primaire et des métastases serait un atout majeur pour le diagnostic précoce et la thérapie des cancers. Dans une double optique de diagnostic et de thérapie, les nanoparticules sont apparues comme des candidats prometteurs pour la mise en place de protocoles thérapeutiques incluant le ciblage de la tumeur, son diagnostic par des techniques d'imagerie optique, ainsi que sa thérapie. Dans ce contexte notre laboratoire développe des appareils d'imagerie optique du petit animal (2D et 3D), afin de permettre le suivi de la biodistribution à long terme et de manière non invasive de sondes fluorescentes et de nanoparticules in vivo. La caractérisation du comportement de différentes nanoparticules (nanocapsules lipidiques, nanoparticules d'oxyde de Gadolinium et nanoparticules d'or) présentant différentes fonctionnalisations de surface a été réalisée in vitro et in vivo sur différents modèles tumoraux. Les résultats obtenus montrent l'influence de la fonctionnalisation et de la taille des nanoparticules sur leur biodistribution in vivo et sur leur voie d'élimination. D'autre part, la fonctionnalisation des nanoparticules par le motif RGD permet le ciblage spécifique des cellules exprimant l'intégrine alpha v beta3 in vitro et améliore l'accumulation tumorale in vivo

Chimio-sensibilisation des cellules tumorales ovariennes par modulation de l'activité de Bcl-xL à l'aide de molécules BH3-mimétiques

CAEN-BU Médecine pharmacie (141182102) / SudocLYON1-BU Santé (693882101) / SudocSudocFranceF

Marqueurs moléculaires et thérapies ciblées (exemple des mutations de l EGFR et de KRAS comme marqueurs prédictifs de réponse aux agents ciblant l EGFR)

Les cancers broncho-pulmonaires (CBNPC) et colorectaux sont les deux premières causes de mortalité par cancer dans le monde. Détectés tardivement, ils sont de très mauvais pronostic, avec une survie à 5 ans respectivement de 12 % et 5 %. Récemment, de nouvelles thérapies dites ciblées, comme par exemple les anticorps monoclonaux anti-EGFR ou les inhibiteurs de l activité tyrosine kinase (TKI) de l EGFR, ont été développées. Une faible proportion des patients porteurs d un cancer broncho-pulmonaire ou colorectal répond à ces traitements ; leur survie est augmentée et leur qualité de vie améliorée. Différentes études ont permis de préciser les facteurs prédictifs de réponse, tels que les mutations des exons 18 à 21 de l EGFR et celles de l exon 1 de KRAS. Ainsi, les anticorps monoclonaux sont indiqués dans le traitement du cancer colorectal métastatique lorsque KRAS est sauvage. Pour le traitement des CBNPC par TKIs, le statut mutationnel de l EGFR est en cours d évaluation. Au laboratoire, l analyse des mutations a dans un premier temps été réalisée par la méthode de référence, le séquençage direct, puis une technique alternative, le pyroséquençage a été développée. Cette méthode est plus sensible, plus rapide et moins coûteuse, et permet une prise en charge optimale des patients pouvant bénéficier d une thérapie ciblant l EGFR.GRENOBLE1-BU Médecine pharm. (385162101) / SudocSudocFranceF

Vectorisation de biomolécules pour l'imagerie et la thérapie des cancers

Le diagnostic précis et précoce des tumeurs, ainsi que le développement de thérapies ciblées sont deux axes principaux de recherche en cancérologie. La génération de vecteurs spécifiques capables d'adresser un agent de contraste et/ou un médicament au niveau de la tumeur primaire et des métastases serait un atout majeur pour le diagnostic précoce et la thérapie des cancers. Dans une double optique de diagnostic et de thérapie, les nanoparticules sont apparues comme des candidats prometteurs pour la mise en place de protocoles thérapeutiques incluant le ciblage de la tumeur, son diagnostic par des techniques d'imagerie optique, ainsi que sa thérapie. Dans ce contexte notre laboratoire développe des appareils d'imagerie optique du petit animal (2D et 3D), afin de permettre le suivi de la biodistribution à long terme et de manière non invasive de sondes fluorescentes et de nanoparticules in vivo.La caractérisation du comportement de différentes nanoparticules (nanocapsules lipidiques, nanoparticules d'oxyde de Gadolinium et nanoparticules d'or) présentant différentes fonctionnalisations de surface a été réalisée in vitro et in vivo sur différents modèles tumoraux. Les résultats obtenus montrent l'influence de la fonctionnalisation et de la taille des nanoparticules sur leur biodistribution in vivo et sur leur voie d'élimination. D'autre part, la fonctionnalisation des nanoparticules par le motif RGD permet le ciblage spécifique des cellules exprimant l'intégrine avP3 in vitro et améliore l'accumulation tumorale in vivo.Early and accurate detection oftumors, as weil as the development oftargeted-treatments are major field ofresearch in oncology. The generation of specifie vectors, capable of addressing a drug or a contrast agent to the primary tumor site as weil as in the remote (micro-) metastasis would be thus a major asset for early diagnosis and cancer therapy. ln this double goal of diagnosis and therapy, nanopartic1es emerged as promising candidates for the establishment of therapeutic protocols, inc1uding targeting, diagnosis with optical imaging techniques and cancer therapy. ln this context, our laboratory developed 2D and 3D optical imaging devices, designed for small animal imaging, to enable the long term non-invasive follow-up of the in vivo biodistribution of fluorescent probes and nanoparticles. The characterization ofthe behavior of different nanopartic1es (lipidic nanocapsules, gadolinium oxide nanopartic1es and gold nanopartic1es) with various surface functionalizations was carried out in vitro and in vivo on different tumor models. The results show the influence of the functionalization and the size-ofnanopartic1es on their in vivo biodistribution and on their way of elimination. On the other hand, the functionalization of the nanopartic1es by the RGD motif allows a specifie targeting of cells expressing the av~3 integrin in vitro and improves the in vivo accumulation in tumors.GRENOBLE1-BU Sciences (384212103) / SudocSudocFranceF

Pyrosequencing, a method approved to detect the two major EGFR mutations for anti EGFR therapy in NSCLC.

International audienceABSTRACT: BACKGROUND: Epidermal Growth Factor Receptor (EGFR) mutations, especially in-frame deletions in exon 19 (delta LRE) and a point mutation in exon 21 (L858R) predict gefitinib sensitivity in patients with non-small cell lung cancer. Several methods are currently described for their detection but the gold standard for tissue samples remains direct DNA sequencing, which requires samples containing at least 50% of tumor cells. METHODS: We designed a pyrosequencing assay based on nested PCR for the characterization of theses mutations on formalin-fixed and paraffin-embedded tumor tissue. RESULTS: This method is highly specific and permits precise characterization of all the exon 19 deletions. Its sensitivity is higher than that of "BigDye terminator" sequencing and enabled detection of 3 additional mutations in the 58 NSCLC tested. The concordance between the two methods was very good (97.4%). In the prospective analysis of 213 samples, 7 (3.3%) samples were not analyzed and EGFR mutations were detected in 18 (8.7%) patients. However, we observed a deficit of mutation detection when the samples were very poor in tumor cells. CONCLUSIONS: pyrosequencing is then a highly accurate method for detecting delta LRE and L858R EGFR mutations in patients with NSCLC when the samples contain at least 20% of tumor cells

Optical small animal imaging in the drug discovery process.

International audienceMolecular imaging of tumors in preclinical models is of the utmost importance for developing innovative cancer treatments. This field is moving extremely rapidly, with recent advances in optical imaging technologies and sophisticated molecular probes for in vivo imaging. The aim of this review is to provide a succinct overview of the imaging modalities available for rodents and with focus on describing optical probes for cancer imaging

EPR-mediated tumor targeting using ultrasmall-hybrid nanoparticles: From animal to human with theranostic AGuIX nanoparticles

International audienceInterest of tumor targeting through EPR effect is still controversial due to intrinsic low targeting efficacy and rare translation to human cancers. Moreover, due to different reasons, it has generally been described for relatively large nanoparticles (NPs) (hydrodynamic diameter > 10 nm). In this review EPR effect will be discussed for ultrasmall NPs using the example of the AGuIX® NP (Activation and Guiding of Irradiation by X-ray) recently translated in clinic. AGuIX® NP is a 4 ± 2 nm hydrodynamic diameter polysiloxane based NP. Since AGuIX® NP biodistribution is monitored by magnetic resonance imaging (MRI) and its activation is triggered by irradiation upon X-rays, this NP is well adapted for a theranostic approach of radiotherapy cancer treatment. Here we show that AGuIX® NP is particularly well suited to benefit from EPR-mediated tumor targeting thanks to an ultrasmall size and efficacy under irradiation at small dose. Indeed, intravenously-injected AGuIX® NP into rodent cancer models passively reached the tumor and revealed no toxicity, favoured by renal clearance. Moreover, translation of AGuIX® NP accumulation and retention into humans carrying brain metastases was validated during a first-in-man phase Ib trial taking advantage of easy biodistribution monitoring by MRI

Aerosol administration of theranostic nanoparticles : from imaging to therapy

International audienc