Epithelial ovarian cancer (EOC) is the most lethal gynecologic cancer and urgently requires new therapies. Oncolytic viruses (OV) are a strong contender. OVs interact with immune components of the TME, which can be altered due to specific genetic mutations. The present study evaluates the impact of specific tumour mutations on the response to carboplatin, the current standard of care, and VSV∆M51, a promising OV candidate. After a study of genetically diverse models, constitutive KRas activation enhanced VSV∆M51 replication in-vitro and sensitivity in syngeneic in-vivo models. VSV∆M51 prolonged survival in syngeneic tumour- bearing mice with KRas, Trp53 and Pten mutations, including one tumour model that did not respond to carboplatin. Response to VSV∆M51 in-vivo was associated with activation of CD4+ and CD8+ T lymphocytes in the peritoneal TME. In summary, VSV∆M51-based immunotherapy has shown promise in diverse murine models of EOC bearing clinically relevant mutations.

Matrix metalloproteinases (MMPs) are a family of zinc endopeptidases capable of digesting the extracellular matrix (ECM), which is essential for tissue structure and transmitting messages between cells. MMPs play an important role in cancer, controlling cell migration, proliferation, apoptosis, regulation of tumour expansion, angiogenesis and invasion. Previous research has indicated high expression of MT1-MMP in breast cancers suggesting a potential role in tumour progression. Our results confirm that 3D multicellular tumour spheroids (MCTS) using phenotype-specific breast cancer cell lines are a valuable experimental model of the tumour microenvironment. Optimisation of MCTS culture growth conditions using different breast cancer cell lines (MCF-7, T47D, MDA-MB-468 and MDA-MB-231) was performed. Unexpected detection of MT1-MMP in MCF-7 MCTS warranted further investigation. MT1-MMP expression in different micro-environmental conditions, including hypoxia and nutrient deprivation (serum-free induced autophagy) were measured in MCF-7 monolayer cultures and MCTS models using immunofluorescence (IF), immunohistochemistry (IHC) and western blot (WB). MT1-MMP expression was rapidly and irreversibly up-regulated in MCF-7 breast cancer cells under conditions of stress (hypoxia and autophagy) compared to normal conditions suggesting an important role of the culture environment on cells behaviour and protein expression. We employed isobaric tags for relative and absolute quantitation (iTRAQ) technology to correlate MT1-MMP increase with proteomic profiles in MCF-7 breast cancer cell grown under hypoxic, serum-free and 3D MCTS conditions. More than 3500 proteins were identified, which were clustered into groups based on response to unique or shared microenvironment changes. Hypoxic monolayer and spheroid cells exhibited changes in anaerobic metabolism and lipid synthesis, respectively, whereas autophagy resulted in up-regulation of cellular component disassembly. The result indicated multiple drivers of MT1-MMP expression in MCF-7 cells. / Al-Mstansiriya University, Iraq

The tumour-associated stroma, consisting of fibroblasts, inflammatory cells, vasculature and extracellular matrix proteins, plays a critical role in tumour growth, but how it regulates cancer cell migration and metastasis is poorly understood. The Wnt-planar cell polarity (PCP) pathway regulates convergent extension movements in vertebrate development. However, it is unclear whether this pathway also functions in cancer cell migration. In addition, the factors that mobilize long-range signalling of Wnt morphogens, which are tightly associated with the plasma membrane, have yet to be completely characterized. Here, I show that fibroblasts secrete membrane microvesicles of endocytic origin, termed exosomes, which promote tumour cell protrusive activity, motility and metastasis via the exosome component Cd81. In addition, I demonstrate that fibroblast exosomes activate autocrine Wnt-PCP signalling in breast cancer cells as detected by the association of Wnt with Fzd receptors and the asymmetric distribution of Fzd-Dvl and Vangl-Pk complexes in exosome-stimulated cancer cell protrusive structures. Moreover, I show that Pk expression in breast cancer cells is essential for fibroblast-stimulated cancer cell metastasis. Lastly, I reveal that trafficking in cancer cells promotes tethering of autocrine Wnt11 to fibroblast exosomes. These studies further our understanding of the role of the tumour-associated stroma in cancer metastasis and bring us closer to a more targeted approach for the treatment of cancer spread.

The tumour-associated stroma, consisting of fibroblasts, inflammatory cells, vasculature and extracellular matrix proteins, plays a critical role in tumour growth, but how it regulates cancer cell migration and metastasis is poorly understood. The Wnt-planar cell polarity (PCP) pathway regulates convergent extension movements in vertebrate development. However, it is unclear whether this pathway also functions in cancer cell migration. In addition, the factors that mobilize long-range signalling of Wnt morphogens, which are tightly associated with the plasma membrane, have yet to be completely characterized. Here, I show that fibroblasts secrete membrane microvesicles of endocytic origin, termed exosomes, which promote tumour cell protrusive activity, motility and metastasis via the exosome component Cd81. In addition, I demonstrate that fibroblast exosomes activate autocrine Wnt-PCP signalling in breast cancer cells as detected by the association of Wnt with Fzd receptors and the asymmetric distribution of Fzd-Dvl and Vangl-Pk complexes in exosome-stimulated cancer cell protrusive structures. Moreover, I show that Pk expression in breast cancer cells is essential for fibroblast-stimulated cancer cell metastasis. Lastly, I reveal that trafficking in cancer cells promotes tethering of autocrine Wnt11 to fibroblast exosomes. These studies further our understanding of the role of the tumour-associated stroma in cancer metastasis and bring us closer to a more targeted approach for the treatment of cancer spread.

Il existe une littérature abondante sur les mécanismes cellulaires de résistance à la chimiothérapie, décrivant notamment les pompes d’efflux, les modifications des cibles (comme les topoisomérases) ou les altérations de l’apoptose. Peu de publications s’intéressent aux mécanismes de chimiorésistance liée au microenvironnement tumoral. Les agents anticancéreux doivent traverser l’interstitium tumoral pour atteindre toutes les cellules (dont les cellules hypoxiques éloignées des vaisseaux sanguins) à des concentrations suffisantes pour être létales. Les modèles de culture cellulaire en couches multiples ont permis de montrer la faible pénétration des molécules de chimiothérapie. Les techniques d’immunohistochimie permettent une mesure quantitative de la distribution de ces molécules à partir des vaisseaux sanguins. Nous avons évalué la pénétration de plusieurs inhibiteurs de topoisomérases : topotécan, doxorubicine, mitoxantrone et banoxantrone. Nous avons comparé la distribution de ces molécules à travers des tissus sains et des tissus tumoraux, démontrant la pénétration limitée des molécules de chimiothérapie dans les tumeurs. Par contre, nous avons montré que la banoxantrone pénètre rapidement et de façon uniforme. Cette pro-drogue est convertit en AQ4 (un inhibiteur de topoisomérase II ressemblant à la mitoxantrone) en condition d’hypoxie. La mitoxantrone cible les cellules bien oxygénées et AQ4 cible les cellules hypoxiques. Cette combinaison de traitement aboutit à une distribution intratumorale complémentaire et à une amélioration de l’activité antitumorale. Ainsi, optimiser la pénétration des chimiothérapies et/ou cibler spécifiquement les cellules hypoxiques peut contourner la chimiorésistance liée au microenvironnement tumoral. / There is a vast literature about mechanisms that lead to drug resistance of individual cancer cells, including drug export pumps, changes in expression of targets (such as topoisomerases) or alterations in apoptosis. A smaller number of publications has drawn attention to causes of drug resistance that depend on the solid tumour microenvironment. Drugs must penetrate the extra-vascular space to reach all of the cancer cells (including cells far from blood vessels in hypoxic condition) in sufficient concentration to cause lethal toxicity. Model systems such as multilayered cell cultures provide direct evidence of poor drug penetration through tumour tissue. In vivo techniques using quantitative immunohistochemistry allow studying drug distribution as a function of distance from the nearest blood vessel. We have evaluated the penetration of several topoisomerase inhibitors: topotecan, doxorubicine, mitoxantrone and banoxantrone (AQ4N). We have compared the distribution of these drugs through normal and tumour tissue, demonstrating the limited perivascular distribution of conventional chemotherapies in tumour. We have also showed the rapid and uniform penetration of banoxantrone. This pro-drug is reduced to AQ4 (a topoisomérase II inhibitor of similar structure to mitoxantrone) under hypoxic condition. The targeting of mitoxantrone to oxygenated regions and AQ4 to hypoxic tumour regions resulted in effective drug exposure over the entire tumour and increased tumour growth delay compared with either drug alone. Improving drug penetration and/or targeting hypoxic tumour cells may overcome chemoresistance due to the tumour microenvironment.

The tumour microenvironment comprises the non-cancerous cells present in the tumour mass (fibroblasts, endothelial, and immune cells), as well as signalling molecules and extracellular matrix. Tumour growth, invasion, metastasis, and response to therapy are influenced by the tumour microenvironment. Therefore, characterising the cellular and molecular components of the tumour microenvironment, and understanding how they influence tumour progression, represent a crucial aim for the success of cancer therapies. High-grade serous ovarian cancer provides an excellent opportunity to systematically study the tumour microenvironment due to its clinical presentation of advanced disseminated disease and debulking surgery being standard of care. This thesis first presents a case report of a long-term survivor (>10 years) of metastatic high-grade serous ovarian cancer who exhibited concomitant regression/progression of the metastatic lesions (5 samples). We found that progressing metastases were characterized by immune cell exclusion, whereas regressing metastases were infiltrated by CD8+ and CD4+ T cells. Through a T cell - neoepitope challenge assay we demonstrated that pre- dicted neoepitopes were recognised by the CD8+ T cells obtained from blood drawn from the patient, suggesting that regressing tumours were subjected to immune attack. Immune excluded tumours presented a higher expression of immunosuppressive Wnt signalling, while infiltrated tumours showed a higher expression of the T cell chemoattractant CXCL9 and evidence of immunoediting. These findings suggest that multiple distinct tumour immune microenvironments can co-exist within a single individual and may explain in part the hetero- geneous fates of metastatic lesions often observed in the clinic post-therapy. Second, this thesis explores the prevalence of intra-patient tumour microenvironment het- erogeneity in high-grade serous ovarian cancer at diagnosis (38 samples from 8 patients), as well as the effect of chemotherapy on the tumour microenvironment (80 paired samples from 40 patients). Whole transcriptome analysis and image-based quantification of T cells from treatment-naive tumours revealed highly prevalent variability in immune signalling and distinct immune microenvironments co-existing within the same individuals at diagnosis. ConsensusTME, a method that generates consensus immune and stromal cell gene signatures by intersecting state-of-the-art deconvolution methods that predict immune cell populations using bulk RNA data was developed. ConsensusTME improved accuracy and sensitivity of T cell and leukocyte deconvolutions in ovarian cancer samples. As previously observed in the case report, Wnt signalling expression positively correlated with immune cell exclusion. To evaluate the effect of chemotherapy on the tumour microenvironment, we compared site-matched and site-unmatched tumours before and after neoadjuvant chemotherapy. Site- matched samples showed increased cytotoxic immune activation and oligoclonal expansion of T cells after chemotherapy, unlike site-unmatched samples where heterogeneity could not be accounted for. In addition, low levels of immune activation pre-chemotherapy were found to be correlated with immune activation upon chemotherapy treatment. These results cor- roborate that the tumour-immune interface in advanced high-grade serous ovarian cancer is intrinsically heterogeneous, and that chemotherapy induces an immunogenic effect mediated by cytotoxic cells. Finally, the different deconvolution methods were benchmarked along with ConsensusTME in a pan-cancer setting by comparing deconvolution scores to DNA-based purity scores, leukocyte methylation data, and tumour infiltrating lymphocyte counts from image analysis. In so far as it has been benchmarked, unlike the other methods, ConsensusTME performs consistently among the top three methods across cancer-related benchmarks. Additionally, ConsensusTME provides a dynamic and evolvable framework that can integrate newer de- convolution tools and benchmark their performance against itself, thus generating an ever updated version. Overall, this thesis presents a systematic characterisation of the tumour microenvironment of high grade serous ovarian cancer in treatment-naive and chemotherapy treated samples, and puts forward the development of an integrative computational method for the systematic analysis of the tumour microenvironment of different tumour types using bulk RNA data.

Orientador: Jose Andres Yunes / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-10T05:42:02Z (GMT). No. of bitstreams: 1 Laranjeira_AngeloBrunelliAlbertoni_M.pdf: 2767287 bytes, checksum: 8b66a1e06cad088fa256103c24e955a7 (MD5) Previous issue date: 2007 / Resumo: A resistência intrínseca ou adquirida aos compostos quimioterápicos é uma das mais importantes causas dos insucessos no tratamento das LLAs pediátricas. A interação da LLA com o microambiente da medula óssea contribui para a proliferação e resistência ao regime quimioterápico das células leucêmicas através de uma grande variedade de mecanismos celulares que provavelmente incluem: aumento da expressão de transportadores celulares, aumento no processo de reparo do DNA, diminuição na regulação dos alvos das drogas, mudanças na regulação do ciclo celular e alteração nas vias apoptóticas. No presente estudo observou-se que a interação estabelecida entre células estromais e células de LLA-B, promoveu a ativação destas como avaliado pela análise das moléculas de superfície das células leucêmicas ao longo dos períodos de cultivo, além da sobrevivência e/ou proliferação em mais de 60% dos casos in vitro. A comunicação entre os dois tipos celulares também mostrou a influência do estroma na modulação da expressão transcricional de 17 genes relacionados com a resistência e sensibilidade a quimioterápicos em células de LLA-B. A modulação teve como conseqüência o aumento nos níveis de expressão da maioria dos genes de resistência e a queda de expressão da maioria dos genes de sensibilidade. Sendo assim, a LLA, pela interação com as células estromais, apresentaram uma alteração que as levou a um fenótipo característico de células resistentes. Essa alteração de expressão mediada pelo contato com o estroma foi confirmada por estudos funcionais de dois genes relacionados com a resistência. O gene KCNN4 em linhagens celulares, que quando submetidas à ação do clotrimazol apresentaram maior viabilidade na presença do que na ausência do estroma; e a adição da proteína recombinante IGFBP-7 no sistema de co-cultura promoveu a resistência e até mesmo proliferação na presença da L-asparaginase. Esta proteína também se mostrou atuante na proliferação das células estromais. Estes resultados mostram dois genes de LLA, que quando modulados pelo contato com o estroma podem contribuir com a maior resistência ao regime quimioterápico, podendo vir a ser usados como alvo para posteriores terapias / Abstract: The intrinsic or acquired chemotherapy resistance composites one of the most important causes of failures in the treatment of pediatric ALL. The ALL and bone marrow microenvironment nteraction contributes for the proliferation and resistance to the chemotherapy regimen of leukemic cells probably through a great variety of cellular mechanisms, including increase of the expression of cellular transporters, increase in the process of DNA repair, downregulation of drugs targets, changes in the regulation of cellular cycle and alteration in the apoptotic ways. In the present study it was observed that the interaction established between stromal cells and pre-B ALL, evaluated through analysis of surface molecules in leukemic cells throughout the periods of culture, were important for the survival and/or proliferation in more than 50% of the cases in vitro. This interaction also showed the influence of stroma in the transcriptional profile of 17 genes related with the resistance and sensitivity to chemotherapeutic agents in pre-B ALL cells. The modulation had as consequence the increase in the levels of expression of the majority of the resistance genes and the decrease of expression of the majority of the sensitivity genes. Being thus, these cellular types, for the interaction with the stromal cells, had presented an alteration that took them to one phenotype characteristic of resistant cells. This stroma-mediated alteration was confirmed by functional studies of two genes related with the resistance. Gene KCNN4 three leukemic cell lines, that when submitted to the action of clotrimazole they had presented greater viability in the presence than in the absence of stroma; and the addition of recombinant protein IGFBP-7 in the co-culture system promoted the resistance and proliferation of primary ALL cells in the presence of the L-asparaginase. This protein also induced proliferation of stromal cells. These results show two genes of ALL, that when modulated for the contact with stroma, can contribute with a resistance to the chemotherapic regimen, becoming possible targets for posterior therapies / Mestrado / Genetica Animal e Evolução / Mestre em Genética e Biologia Molecular

Background To investigate vascular-related pathophysiological characteristics of two human lung cancers with modifiable vascularisation using MRI and CT. Methods Tumour xenografts with modifiable vascularisation were established in 71 rats (approval by the Animal Care Committee was obtained) by subcutaneous transplantation of two human non-small-cell lung cancer (NSCLC) cells (A549, H1299) either alone or co-transplanted with vascular growth promoters. The vascularity of the tumours was assessed noninvasively by MRI diffusion-weighted-imaging (DWI), T2-weighted, and time-of-flight (TOF) sequences) as well as contrast-enhanced CT (CE-CT), using clinical scanners. As a reference standard, histological examinations (CD-31, fluorescent beads) were done after explantation. Results Microvessel density (MVD) was higher in co-transplanted tumours (171 ± 19 number/mm2) than in non-co-transplanted tumours (111 ± 11 number/mm2; p = 0.002). Co-transplanted tumours showed higher growth rates and larger tumour vessels at TOF-MRI as well as larger necrotic areas at CE-CT. In co-transplanted tumours, DWI revealed higher cellularity (lower minimal ADCdiff 166 ± 15 versus 346 ± 27 mm2/s × 10−6; p < 0.001), highly necrotic areas (higher maximal ADCdiff 1695 ± 65 versus 1320 ± 59 mm2/s × 10−6; p < 0.001), and better-perfused tumour stroma (higher ADCperf 723 ± 36 versus 636 ± 51 mm2/s × 10−6; p = 0.005). Significant correlations were found using qualitative and quantitative parameters: maximal ADCperf and MVD (r = 0.326); maximal ADCdiff and relative necrotic volume on CE-CT (r = 0.551); minimal ADCdiff and MVD (r = −0.395). Conclusions Pathophysiological differences related to vascular supply in two human lung cancer cell lines with modifiable vascularity are quantifiable with clinical imaging techniques. Imaging parameters of vascularisation correlated with the results of histology. DWI was able to characterise both the extent of necrosis and the level of perfusion.

Breast cancer is a global public health problem since it is the most frequently diagnosed cancer in women in Western countries. Clinical guidelines for breast cancer prognosis/diagnosis are currently based on tumour size, histological type and grade, lymph node status as well as the expression of various cellular receptors. Yet, current predictions remain unsatisfactory to identify the best treatment for the individual patient. The search for identifying new predictive and prognostic factors is ongoing. Furthermore, compelling evidences have solidified the notion that the evolving epithelial cells, founders of the breast disease, are helped in their malignant course by the tumour microenvironment. Better characterizing the dual effect of the immune regulation but also the epithelial-stromal cross-talk on both tumour-promotion and -suppression is essential for understanding patient uniqueness and their implication in disease outcome. Because of its potential to probe tissues and cells at the molecular level without requirement for extrinsic contrast agents, infrared spectroscopy was seen as an attractive tool for clinical and diagnostic analysis in order to complement the existing methods. <p>In a first step, recording and processing methodology had to be defined in order to optimally compare IR spectra. The methodology developed and the analysis tools tested on carcinoma cell lines, demonstrated that spectra could be distinguished based on the cell line phenotypic nature. <p>The potential of IR imaging for breast tissular structure differentiation was highlighted in this thesis, demonstrating that spectral signature can be correlated with the major histological cell types observed in breast disease tissues. In order to develop a robust algorithm translating spectral data into helpful histopathological information, a spectral database of histologically well-defined breast tissues was built and used for the development of a cell type classifier. This latter one was extensively validated on independent clinical cases. Firstly, the IR-based histopathological classifier correctly assigned spectra acquired on eleven breast disease samples based on their histological nature. Secondly, lymphocyte and Collagen & Fibroblasts spectral signatures were demonstrated to be independent from tissue type and organ since, although trained on reference spectra recorded into breast disease samples, the cell type classifier correctly assigned spectra acquired on lymph nodes/tonsils and scar tissues respectively. Thirdly, we concluded that spectroscopically, breast carcinoma cell lines in culture are well-suited tumour models since spectra acquired on these carcinoma cell lines were correctly recognized as epithelium by the IR-based histological classifier. <p>By spectral characterizing lymphocytes from lymph nodes and tonsils, we demonstrated that the spectra acquired contained enough information to statistically discriminate them according to their lymphocyte activation states. Although considered as activated, the breast disease lymphoid infiltrates were found to present distinct spectral signature from lymphocytes acquired on activated lymph nodes and tonsils. Furthermore, tumour microenvironment, characterized by IR-imaging was demonstrated to exhibit a distinct spectral signature from wound healing tissues. These studies proved the uniqueness of the signature of both lymphoid infiltrate and tumour microenvironment in breast disease context. Correlating these specific spectral signatures to patient outcome and therapeutics response could help better consider the uniqueness of the patient. In a last step, considering the epithelial signature of carcinomas of both low and high grades, we demonstrated that the biochemical information reflected in the IR micro-spectra was clinically relevant for grading purpose.<p><p> <p><p>Le cancer du sein est le cancer le plus fréquemment diagnostiqué chez les femmes dans les pays occidentaux. Jusqu’à peu, les cellules épithéliales tumorales étaient vues comme les seuls acteurs de la carcinogenèse ;processus se déroulant dans un milieu extracellulaire considéré au pire comme passif ou permissif à l’évolution tumorale des cellules épithéliales adjacentes. Cependant, de nombreuses études ont montré que ce microenvironnement tumoral pouvait soit promouvoir le processus de carcinogenèse soit le combattre empêchant par la même, l’occurrence de la maladie. <p>Ce projet de thèse s’inscrit dans une problématique actuelle, à savoir une meilleure compréhension de la maladie mais également une prise en charge plus individualisée des patientes. Nous abordons ici une voie de recherche novatrice basée sur la signature globale des molécules cellulaires via leur spectre infrarouge. La technologie utilisée, à savoir la spectroscopie infrarouge, nous fournit une observation quantitative et qualitative de milliers de vibrations moléculaires. L’adaptation de réseaux de plusieurs milliers de détecteurs indépendants aux microscopes infrarouges permet, grâce aux méthodes statistiques multivariées, d’investiguer l’architecture macromoléculaire des cellules au sein d’une coupe tissulaire et de corréler les informations spectrales ainsi obtenues à l’histopathologie des tissus. Par cette technologie, nous visons à mettre au point un outil diagnostique et pronostique pour le cancer du sein basé sur l’imagerie IR. <p>Durant ce projet, nous avons montré que les différents types cellulaires observés dans les carcinomes mammaires pouvaient être distingués par le biais de leur spectre IR, qu’un modèle de reconnaissance histologique pouvait être construit, validé et surtout automatisé et que ce modèle pouvait être transposé à l’étude d’autres tissus (ganglions, amygdales et cicatrices) et d’autres types d’échantillons (cellules épithéliales en culture). Nous avons également montré que les spectres de cellules épithéliales pouvaient être corrélés au grade histopathologique de la tumeur. Les spectres acquis de ganglions/amygdales ont montré que les profils spectraux pouvaient être corrélés à l’état d’activation lymphocytaire. De plus, l’étude de l’état d’activation lymphocytaire et fibroblastique a permis de mettre en avant un profil spectral propre et bien distinct des infiltrats lymphocytaires d’une part et de la matrice extracellulaire aux abords des tumeurs invasives d’autre part. <p> / Doctorat en Sciences agronomiques et ingénierie biologique / info:eu-repo/semantics/nonPublished

Background To investigate vascular-related pathophysiological characteristics of two human lung cancers with modifiable vascularisation using MRI and CT. Methods Tumour xenografts with modifiable vascularisation were established in 71 rats (approval by the Animal Care Committee was obtained) by subcutaneous transplantation of two human non-small-cell lung cancer (NSCLC) cells (A549, H1299) either alone or co-transplanted with vascular growth promoters. The vascularity of the tumours was assessed noninvasively by MRI diffusion-weighted-imaging (DWI), T2-weighted, and time-of-flight (TOF) sequences) as well as contrast-enhanced CT (CE-CT), using clinical scanners. As a reference standard, histological examinations (CD-31, fluorescent beads) were done after explantation. Results Microvessel density (MVD) was higher in co-transplanted tumours (171 ± 19 number/mm2) than in non-co-transplanted tumours (111 ± 11 number/mm2; p = 0.002). Co-transplanted tumours showed higher growth rates and larger tumour vessels at TOF-MRI as well as larger necrotic areas at CE-CT. In co-transplanted tumours, DWI revealed higher cellularity (lower minimal ADCdiff 166 ± 15 versus 346 ± 27 mm2/s × 10−6; p < 0.001), highly necrotic areas (higher maximal ADCdiff 1695 ± 65 versus 1320 ± 59 mm2/s × 10−6; p < 0.001), and better-perfused tumour stroma (higher ADCperf 723 ± 36 versus 636 ± 51 mm2/s × 10−6; p = 0.005). Significant correlations were found using qualitative and quantitative parameters: maximal ADCperf and MVD (r = 0.326); maximal ADCdiff and relative necrotic volume on CE-CT (r = 0.551); minimal ADCdiff and MVD (r = −0.395). Conclusions Pathophysiological differences related to vascular supply in two human lung cancer cell lines with modifiable vascularity are quantifiable with clinical imaging techniques. Imaging parameters of vascularisation correlated with the results of histology. DWI was able to characterise both the extent of necrosis and the level of perfusion.