The Experts below are selected from a list of 360 Experts worldwide ranked by ideXlab platform
Donna M. Peehl - One of the best experts on this subject based on the ideXlab platform.
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bioorthogonal labeling of human prostate cancer Tissue Slice cultures for glycoproteomics
Angewandte Chemie, 2017Co-Authors: David R Spiciarich, Rosalie Nolley, Donna M. Peehl, Sophia L Maund, Sean C Purcell, Jason Herschel, Anthony T Iavarone, Carolyn R BertozziAbstract:Sialylated glycans are found at elevated levels in many types of cancer and have been implicated in disease progression. However, the specific glycoproteins that contribute to the cancer cell-surface sialylation are not well characterized, specifically in bona fide human disease Tissue. Metabolic and bioorthogonal labeling methods have previously enabled the enrichment and identification of sialoglycoproteins from cultured cells and model organisms. Herein, we report the first application of this glycoproteomic platform to human Tissues cultured ex vivo. Both normal and cancerous prostate Tissues were Sliced and cultured in the presence of the azide-functionalized sialic acid biosynthetic precursor Ac4ManNAz. The compound was metabolized to the azidosialic acid and incorporated into cell surface and secreted sialoglycoproteins. Chemical biotinylation followed by enrichment and mass spectrometry led to the identification of glycoproteins that were found at elevated levels or uniquely in cancerous prostate Tissue. This work therefore extends the use of bioorthogonal labeling strategies to problems of clinical relevance.
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a Tissue graft model of dna damage response in the normal and malignant human prostate
The Journal of Urology, 2014Co-Authors: Taija Af M Hallstrom, Rosalie Nolley, Junqiang Tian, Marikki Laiho, Stephen Reese, Ville Rantanen, Hongjuan Zhao, Donna M. PeehlAbstract:Purpose: DNA damage responses are relevant to prostate cancer initiation, progression and treatment. Few models of the normal and malignant human prostate that maintain stromal-epithelial interactions in vivo exist in which to study DNA damage responses. We evaluated the feasibility of maintaining Tissue Slice grafts at subcutaneous vs subrenal capsular sites in RAG2−/−γC−/− mice to study the DNA damage responses of normal and malignant glands.Materials and Methods: We compared the take rate and histology of Tissue Slice grafts from fresh, precision cut surgical specimens that were maintained for 1 to 4 weeks in subcutaneous vs subrenal capsular sites. Induction of γH2AX, p53, ATM and apoptosis was evaluated as a measure of the DNA damage response after irradiation.Results: The take rate of subcutaneous Tissue Slice grafts was higher than typically reported but lower than at the subrenal capsular site. Subcutaneous Tissue Slice grafts frequently showed basal cell hyperplasia, squamous metaplasia and cysti...
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Tissue Slice grafts of human renal cell carcinoma an authentic preclinical model with high engraftment rate and metastatic potential
Urologic Oncology-seminars and Original Investigations, 2014Co-Authors: Alan Thong, Rosalie Nolley, Hongjuan Zhao, Jennifer Santos, Alexandre Ingels, Maija Valta, Sarah R Young, Donna M. PeehlAbstract:Objective Discovery of curative therapies for renal cell carcinoma (RCC) is hampered by lack of authentic preclinical models. Tumorgrafts, generated by direct implantation of patient-derived Tissues into mice, have demonstrated superior ability to predict therapeutic response. We evaluated “Tissue Slice grafts” (TSGs) as an improved tumorgraft model of RCC.
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patient derived Tissue Slice grafts accurately depict response of high risk primary prostate cancer to androgen deprivation therapy
Journal of Translational Medicine, 2013Co-Authors: Hongjuan Zhao, Rosalie Nolley, Stephen Reese, Jennifer Santos, Alexandre Ingels, Alan Thong, Donna M. PeehlAbstract:Effective eradication of high-risk primary prostate cancer (HRPCa) could significantly decrease mortality from prostate cancer. However, the discovery of curative therapies for HRPCa is hampered by the lack of authentic preclinical models. We improved upon tumorgraft models that have been shown to predict drug response in other cancer types by implanting thin, precision-cut Slices of HRPCa under the renal capsule of immunodeficient mice. Tissue Slice grafts (TSGs) from 6 cases of HRPCa were established in mice. Following androgen deprivation by castration, TSGs were recovered and the presence and phenotype of cancer cells were evaluated. High-grade cancer in TSGs generated from HRPCa displayed characteristic Gleason patterns and biomarker expression. Response to androgen deprivation therapy (ADT) was as in humans, with some cases exhibiting complete pathologic regression and others showing resistance to castration. As in humans, ADT decreased cell proliferation and prostate-specific antigen expression in TSGs. Adverse pathological features of parent HRPCa were associated with lack of regression of cancer in corresponding TSGs after ADT. Castration-resistant cancer cells remaining in TSGs showed upregulated expression of androgen receptor target genes, as occurs in castration-resistant prostate cancer (CRPC) in humans. Finally, a rare subset of castration-resistant cancer cells in TSGs underwent epithelial-mesenchymal transition, a process also observed in CRPC in humans. Our study demonstrates the feasibility of generating TSGs from multiple patients and of generating a relatively large number of TSGs from the same HRPCa specimen with similar cell composition and histology among control and experimental samples in an in vivo setting. The authentic response of TSGs to ADT, which has been extensively characterized in humans, suggests that TSGs can serve as a surrogate model for clinical trials to achieve rapid and less expensive screening of therapeutics for HRPCa and primary CRPC.
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metabolic reprogramming and validation of hyperpolarized 13c lactate as a prostate cancer biomarker using a human prostate Tissue Slice culture bioreactor
The Prostate, 2013Co-Authors: Kayvan R Keshari, Donna M. Peehl, Renuka Sriram, Mark Van Criekinge, David M Wilson, Zhen J Wang, Daniel B Vigneron, John KurhanewiczAbstract:BACKGROUND The treatment of prostate cancer has been impeded by the lack of both clinically relevant disease models and metabolic markers that track tumor progression. Hyperpolarized (HP) 13C MR spectroscopy has emerged as a new technology to investigate the metabolic shifts in prostate cancer. In this study, we investigate the glucose reprogramming using HP 13C pyruvate MR in a patient-derived prostate Tissue Slice culture (TSC) model. METHODS The steady-state metabolite concentrations in freshly excised human prostate TSCs were assessed and compared to those from snap-frozen biopsy samples. The TSCs were then applied to a perfused cell (bioreactor) platform, and the bioenergetics and the dynamic pyruvate flux of the TSCs were investigated by 31P and HP 13C MR, respectively. RESULTS The prostate TSCs demonstrated steady-state glycolytic and phospholipid metabolism, and bioenergetics that recapitulate features of prostate cancer in vivo. 13C spectra following injection of HP 13C pyruvate showed significantly increased pyruvate to lactate flux in malignant as compared to the benign prostate TSCs. This increased flux in the malignant prostate TSCs correlated with both increased expression of monocarboxylate transporters (MCT) and activity of lactate dehydrogenase (LDH). CONCLUSIONS We provide the first mechanistic evidence for HP 13C lactate as a prostate cancer biomarker in living human Tissues, critical for the interpretation of in vivo studies. More broadly, the clinically relevant metabolic model system in combination with HP MR can facilitate the identification of clinically translatable biomarkers of prostate cancer presence, aggressiveness, and treatment response. Prostate 73: 1171–1181, 2013. © 2013 Wiley Periodicals, Inc.
Rosalie Nolley - One of the best experts on this subject based on the ideXlab platform.
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bioorthogonal labeling of human prostate cancer Tissue Slice cultures for glycoproteomics
Angewandte Chemie, 2017Co-Authors: David R Spiciarich, Rosalie Nolley, Donna M. Peehl, Sophia L Maund, Sean C Purcell, Jason Herschel, Anthony T Iavarone, Carolyn R BertozziAbstract:Sialylated glycans are found at elevated levels in many types of cancer and have been implicated in disease progression. However, the specific glycoproteins that contribute to the cancer cell-surface sialylation are not well characterized, specifically in bona fide human disease Tissue. Metabolic and bioorthogonal labeling methods have previously enabled the enrichment and identification of sialoglycoproteins from cultured cells and model organisms. Herein, we report the first application of this glycoproteomic platform to human Tissues cultured ex vivo. Both normal and cancerous prostate Tissues were Sliced and cultured in the presence of the azide-functionalized sialic acid biosynthetic precursor Ac4ManNAz. The compound was metabolized to the azidosialic acid and incorporated into cell surface and secreted sialoglycoproteins. Chemical biotinylation followed by enrichment and mass spectrometry led to the identification of glycoproteins that were found at elevated levels or uniquely in cancerous prostate Tissue. This work therefore extends the use of bioorthogonal labeling strategies to problems of clinical relevance.
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a Tissue graft model of dna damage response in the normal and malignant human prostate
The Journal of Urology, 2014Co-Authors: Taija Af M Hallstrom, Rosalie Nolley, Junqiang Tian, Marikki Laiho, Stephen Reese, Ville Rantanen, Hongjuan Zhao, Donna M. PeehlAbstract:Purpose: DNA damage responses are relevant to prostate cancer initiation, progression and treatment. Few models of the normal and malignant human prostate that maintain stromal-epithelial interactions in vivo exist in which to study DNA damage responses. We evaluated the feasibility of maintaining Tissue Slice grafts at subcutaneous vs subrenal capsular sites in RAG2−/−γC−/− mice to study the DNA damage responses of normal and malignant glands.Materials and Methods: We compared the take rate and histology of Tissue Slice grafts from fresh, precision cut surgical specimens that were maintained for 1 to 4 weeks in subcutaneous vs subrenal capsular sites. Induction of γH2AX, p53, ATM and apoptosis was evaluated as a measure of the DNA damage response after irradiation.Results: The take rate of subcutaneous Tissue Slice grafts was higher than typically reported but lower than at the subrenal capsular site. Subcutaneous Tissue Slice grafts frequently showed basal cell hyperplasia, squamous metaplasia and cysti...
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Tissue Slice grafts of human renal cell carcinoma an authentic preclinical model with high engraftment rate and metastatic potential
Urologic Oncology-seminars and Original Investigations, 2014Co-Authors: Alan Thong, Rosalie Nolley, Hongjuan Zhao, Jennifer Santos, Alexandre Ingels, Maija Valta, Sarah R Young, Donna M. PeehlAbstract:Objective Discovery of curative therapies for renal cell carcinoma (RCC) is hampered by lack of authentic preclinical models. Tumorgrafts, generated by direct implantation of patient-derived Tissues into mice, have demonstrated superior ability to predict therapeutic response. We evaluated “Tissue Slice grafts” (TSGs) as an improved tumorgraft model of RCC.
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patient derived Tissue Slice grafts accurately depict response of high risk primary prostate cancer to androgen deprivation therapy
Journal of Translational Medicine, 2013Co-Authors: Hongjuan Zhao, Rosalie Nolley, Stephen Reese, Jennifer Santos, Alexandre Ingels, Alan Thong, Donna M. PeehlAbstract:Effective eradication of high-risk primary prostate cancer (HRPCa) could significantly decrease mortality from prostate cancer. However, the discovery of curative therapies for HRPCa is hampered by the lack of authentic preclinical models. We improved upon tumorgraft models that have been shown to predict drug response in other cancer types by implanting thin, precision-cut Slices of HRPCa under the renal capsule of immunodeficient mice. Tissue Slice grafts (TSGs) from 6 cases of HRPCa were established in mice. Following androgen deprivation by castration, TSGs were recovered and the presence and phenotype of cancer cells were evaluated. High-grade cancer in TSGs generated from HRPCa displayed characteristic Gleason patterns and biomarker expression. Response to androgen deprivation therapy (ADT) was as in humans, with some cases exhibiting complete pathologic regression and others showing resistance to castration. As in humans, ADT decreased cell proliferation and prostate-specific antigen expression in TSGs. Adverse pathological features of parent HRPCa were associated with lack of regression of cancer in corresponding TSGs after ADT. Castration-resistant cancer cells remaining in TSGs showed upregulated expression of androgen receptor target genes, as occurs in castration-resistant prostate cancer (CRPC) in humans. Finally, a rare subset of castration-resistant cancer cells in TSGs underwent epithelial-mesenchymal transition, a process also observed in CRPC in humans. Our study demonstrates the feasibility of generating TSGs from multiple patients and of generating a relatively large number of TSGs from the same HRPCa specimen with similar cell composition and histology among control and experimental samples in an in vivo setting. The authentic response of TSGs to ADT, which has been extensively characterized in humans, suggests that TSGs can serve as a surrogate model for clinical trials to achieve rapid and less expensive screening of therapeutics for HRPCa and primary CRPC.
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Abstract B32: Patient-derived Tissue Slice grafts of high-risk primary prostate cancer: An authentic preclinical model for synthetic lethality-based therapy
Connections to Therapy, 2013Co-Authors: Hongjuan Zhao, Rosalie Nolley, Stephen Reese, Alan E. Thong, Jennifer Santos, Alexandre Ingels, Donna M. PeehlAbstract:Background: Effective eradication of high-risk primary prostate cancer (HRPCa) could significantly decrease mortality from prostate cancer. Drugs exploiting synthetic lethality have gained attention as novel anticancer agents. However, the discovery of such therapies for HRPCa is hampered by the lack of authentic preclinical models. Methods: We improved upon tumorgraft models that have been shown to predict drug response in other cancer types by implanting thin, precision-cut Slices of HRPCa under the renal capsule of immunodeficient mice. Tissue Slice grafts (TSGs) from 6 cases of HRPCa were established in mice. Following androgen deprivation by castration, TSGs were recovered and the presence and phenotype of cancer cells were evaluated. Results: High-grade cancer in TSGs generated from HRPCa displayed characteristic Gleason patterns and biomarker expression. Response to androgen deprivation therapy (ADT) was as in humans, with some cases exhibiting complete pathologic regression and others showing resistance to castration. As in humans, ADT decreased cell proliferation and prostate-specific antigen expression in TSGs. Adverse pathological features of parent HRPCa were associated with lack of regression of cancer in corresponding TSGs after ADT. Castration-resistant cancer cells remaining in TSGs showed upregulated expression of androgen receptor target genes, as occurs in castration-resistant prostate cancer (CRPC) in humans. Finally, a rare subset of castration-resistant cancer cells in TSGs underwent epithelial-mesenchymal transition, a process also observed in CRPC in humans. Conclusions: Our study demonstrates the feasibility of generating TSGs from multiple patients and of generating a relatively large number of TSGs from the same HRPCa specimen with similar cell composition and histology among control and experimental samples in an in vivo setting. The authentic response of TSGs to ADT, which has been extensively characterized in humans, suggests that TSGs can serve as a surrogate model for clinical trials to achieve rapid and less expensive screening of synthetic lethality based therapeutics for HRPCa and primary CRPC. Citation Format: Hongjuan Zhao, Alan Thong, Rosie Nolley, Stephen Reese, Jennifer Santos, Alexandre Ingels, Donna Peehl. Patient-derived Tissue Slice grafts of high-risk primary prostate cancer: An authentic preclinical model for synthetic lethality-based therapy. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Synthetic Lethal Approaches to Cancer Vulnerabilities; May 17-20, 2013; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(5 Suppl):Abstract nr B32.
Achim Aigner - One of the best experts on this subject based on the ideXlab platform.
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glioblastoma Tissue Slice tandem cultures for quantitative evaluation of inhibitory effects on invasion and growth
Cancers, 2020Co-Authors: Vasile Sidorcenco, Luisa Krahnen, Marion Schulz, Janina Remy, Donat Kogel, Achim Temme, Ute Krugel, Heike Franke, Achim AignerAbstract:Glioblastomas (GBMs) are the most malignant brain tumors and are essentially incurable even after extensive surgery, radiotherapy, and chemotherapy, mainly because of extensive infiltration of tumor cells into the adjacent normal Tissue. Thus, the evaluation of novel drugs in malignant glioma treatment requires sophisticated ex vivo models that approach the authentic interplay between tumor and host environment while avoiding extensive in vivo studies in animals. This paper describes the standardized setup of an organotypic brain Tissue Slice tandem-culture system, comprising of normal brain Tissue from adult mice and tumor Tissue from human glioblastoma xenografts, and explore its utility for assessing inhibitory effects of test drugs. The microscopic analysis of vertical sections of the Slice tandem-cultures allows for the simultaneous assessment of (i) the invasive potential of single cells or cell aggregates and (ii) the space occupying growth of the bulk tumor mass, both contributing to malignant tumor progression. The comparison of Tissue Slice co-cultures with spheroids vs. Tissue Slice tandem-cultures using tumor xenograft Slices demonstrates advantages of the xenograft tandem approach. The direct and facile application of test drugs is shown to exert inhibitory effects on bulk tumor growth and/or tumor cell invasion, and allows their precise quantitation. In conclusion, we describe a straightforward ex vivo system mimicking the in vivo situation of the tumor mass and the normal brain in GBM patients. It reduces animal studies and allows for the direct and reproducible application of test drugs and the precise quantitation of their effects on the bulk tumor mass and on the tumor's invasive properties.
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Optimized polyethylenimine (PEI)-based nanoparticles for siRNA delivery, analyzed in vitro and in an ex vivo tumor Tissue Slice culture model
Drug Delivery and Translational Research, 2017Co-Authors: Alexander Ewe, Heike Franke, Lea Merz, Sabrina Hobel, Sonja Kallendrusch, Ingo Bechmann, Felicitas Merz, Claudia Heine, Achim AignerAbstract:The non-viral delivery of small RNA molecules like siRNAs still poses a major bottleneck for their successful application in vivo. This is particularly true with regard to crossing physiological barriers upon systemic administration. We have previously established polyethylenimine (PEI)-based complexes for therapeutic RNA formulation. These nanoplexes mediate full RNA protection against nucleolytic degradation, delivery to target Tissues as well as cellular uptake, intracellular release and therapeutic efficacy in preclinical in vivo models. We herein present data on different polyplex modifications for the defined improvement of physicochemical and biological nanoparticle properties and for targeted delivery. (i) By non-covalent modifications of PEI polyplexes with phospholipid liposomes, ternary complexes (“lipopolyplexes”) are obtained that combine the favorable features of PEI and lipid systems. Decreased cytotoxicity and highly efficient delivery of siRNA is achieved. Some lipopolyplexes also allow prolonged storage, thus providing formulations with higher stability. (ii) Novel tyrosine modifications of low molecular weight PEI offer further improvement of stability, biocompatibility, and knockdown efficacy of resulting nanoparticles. (iii) For ligand-mediated uptake, the shielding of surface charges is a critical requirement. This is achieved by PEI grafting with polyethylene glycol (PEG), prior to covalent coupling of anti-HER1 antibodies (Erbitux®) as ligand for targeted delivery and uptake. Beyond tumor cell culture, analyses are extended towards tumor Slice cultures from tumor xenograft Tissues which reflect more realistically the in vivo situation. The determination of siRNA-mediated knockdown of endogenous target genes, i.e., the oncogenic survival factor survivin and the oncogenic receptor tyrosine kinase HER2, reveals nanoparticle penetration and biological efficacy also under intact Tissue and stroma conditions.
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tumor Tissue Slice cultures as a platform for analyzing Tissue penetration and biological activities of nanoparticles
European Journal of Pharmaceutics and Biopharmaceutics, 2017Co-Authors: Lea Merz, Heike Franke, Sabrina Hobel, Sonja Kallendrusch, Alexander Ewe, Ingo Bechmann, Felicitas Merz, Achim AignerAbstract:The success of therapeutic nanoparticles depends, among others, on their ability to penetrate a Tissue for actually reaching the target cells, and their efficient cellular uptake in the context of intact Tissue and stroma. Various nanoparticle modifications have been implemented for altering physicochemical and biological properties. Their analysis, however, so far mainly relies on cell culture experiments which only poorly reflect the in vivo situation, or is based on in vivo experiments that are often complicated by whole-body pharmacokinetics and are rather tedious especially when analyzing larger nanoparticle sets. For the more precise analysis of nanoparticle properties at their desired site of action, efficient ex vivo systems closely mimicking in vivo Tissue properties are needed. In this paper, we describe the setup of organotypic tumor Tissue Slice cultures for the analysis of Tissue-penetrating properties and biological activities of nanoparticles. As a model system, we employ 350μm thick Slice cultures from different tumor xenograft Tissues, and analyze modified or non-modified polyethylenimine (PEI) complexes as well as their lipopolyplex derivatives for siRNA delivery. The described conditions for Tissue Slice preparation and culture ensure excellent Tissue preservation for at least 14days, thus allowing for prolonged experimentation and analysis. When using fluorescently labeled siRNA for complex visualization, fluorescence microscopy of cryo-sectioned Tissue Slices reveals different degrees of nanoparticle Tissue penetration, dependent on their surface charge. More importantly, the determination of siRNA-mediated knockdown efficacies of an endogenous target gene, the oncogenic survival factor Survivin, reveals the possibility to accurately assess biological nanoparticle activities in situ, i.e. in living cells in their original environment. Taken together, we establish tumor (xenograft) Tissue Slices for the accurate and facile ex vivo assessment of important biological nanoparticle properties. Beyond the quantitative analysis of nanoparticle Tissue-penetration, the excellent Tissue preservation and cell viability also allows for the evaluation of biological activities.
Hongjuan Zhao - One of the best experts on this subject based on the ideXlab platform.
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a Tissue graft model of dna damage response in the normal and malignant human prostate
The Journal of Urology, 2014Co-Authors: Taija Af M Hallstrom, Rosalie Nolley, Junqiang Tian, Marikki Laiho, Stephen Reese, Ville Rantanen, Hongjuan Zhao, Donna M. PeehlAbstract:Purpose: DNA damage responses are relevant to prostate cancer initiation, progression and treatment. Few models of the normal and malignant human prostate that maintain stromal-epithelial interactions in vivo exist in which to study DNA damage responses. We evaluated the feasibility of maintaining Tissue Slice grafts at subcutaneous vs subrenal capsular sites in RAG2−/−γC−/− mice to study the DNA damage responses of normal and malignant glands.Materials and Methods: We compared the take rate and histology of Tissue Slice grafts from fresh, precision cut surgical specimens that were maintained for 1 to 4 weeks in subcutaneous vs subrenal capsular sites. Induction of γH2AX, p53, ATM and apoptosis was evaluated as a measure of the DNA damage response after irradiation.Results: The take rate of subcutaneous Tissue Slice grafts was higher than typically reported but lower than at the subrenal capsular site. Subcutaneous Tissue Slice grafts frequently showed basal cell hyperplasia, squamous metaplasia and cysti...
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Tissue Slice grafts of human renal cell carcinoma an authentic preclinical model with high engraftment rate and metastatic potential
Urologic Oncology-seminars and Original Investigations, 2014Co-Authors: Alan Thong, Rosalie Nolley, Hongjuan Zhao, Jennifer Santos, Alexandre Ingels, Maija Valta, Sarah R Young, Donna M. PeehlAbstract:Objective Discovery of curative therapies for renal cell carcinoma (RCC) is hampered by lack of authentic preclinical models. Tumorgrafts, generated by direct implantation of patient-derived Tissues into mice, have demonstrated superior ability to predict therapeutic response. We evaluated “Tissue Slice grafts” (TSGs) as an improved tumorgraft model of RCC.
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patient derived Tissue Slice grafts accurately depict response of high risk primary prostate cancer to androgen deprivation therapy
Journal of Translational Medicine, 2013Co-Authors: Hongjuan Zhao, Rosalie Nolley, Stephen Reese, Jennifer Santos, Alexandre Ingels, Alan Thong, Donna M. PeehlAbstract:Effective eradication of high-risk primary prostate cancer (HRPCa) could significantly decrease mortality from prostate cancer. However, the discovery of curative therapies for HRPCa is hampered by the lack of authentic preclinical models. We improved upon tumorgraft models that have been shown to predict drug response in other cancer types by implanting thin, precision-cut Slices of HRPCa under the renal capsule of immunodeficient mice. Tissue Slice grafts (TSGs) from 6 cases of HRPCa were established in mice. Following androgen deprivation by castration, TSGs were recovered and the presence and phenotype of cancer cells were evaluated. High-grade cancer in TSGs generated from HRPCa displayed characteristic Gleason patterns and biomarker expression. Response to androgen deprivation therapy (ADT) was as in humans, with some cases exhibiting complete pathologic regression and others showing resistance to castration. As in humans, ADT decreased cell proliferation and prostate-specific antigen expression in TSGs. Adverse pathological features of parent HRPCa were associated with lack of regression of cancer in corresponding TSGs after ADT. Castration-resistant cancer cells remaining in TSGs showed upregulated expression of androgen receptor target genes, as occurs in castration-resistant prostate cancer (CRPC) in humans. Finally, a rare subset of castration-resistant cancer cells in TSGs underwent epithelial-mesenchymal transition, a process also observed in CRPC in humans. Our study demonstrates the feasibility of generating TSGs from multiple patients and of generating a relatively large number of TSGs from the same HRPCa specimen with similar cell composition and histology among control and experimental samples in an in vivo setting. The authentic response of TSGs to ADT, which has been extensively characterized in humans, suggests that TSGs can serve as a surrogate model for clinical trials to achieve rapid and less expensive screening of therapeutics for HRPCa and primary CRPC.
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Abstract B32: Patient-derived Tissue Slice grafts of high-risk primary prostate cancer: An authentic preclinical model for synthetic lethality-based therapy
Connections to Therapy, 2013Co-Authors: Hongjuan Zhao, Rosalie Nolley, Stephen Reese, Alan E. Thong, Jennifer Santos, Alexandre Ingels, Donna M. PeehlAbstract:Background: Effective eradication of high-risk primary prostate cancer (HRPCa) could significantly decrease mortality from prostate cancer. Drugs exploiting synthetic lethality have gained attention as novel anticancer agents. However, the discovery of such therapies for HRPCa is hampered by the lack of authentic preclinical models. Methods: We improved upon tumorgraft models that have been shown to predict drug response in other cancer types by implanting thin, precision-cut Slices of HRPCa under the renal capsule of immunodeficient mice. Tissue Slice grafts (TSGs) from 6 cases of HRPCa were established in mice. Following androgen deprivation by castration, TSGs were recovered and the presence and phenotype of cancer cells were evaluated. Results: High-grade cancer in TSGs generated from HRPCa displayed characteristic Gleason patterns and biomarker expression. Response to androgen deprivation therapy (ADT) was as in humans, with some cases exhibiting complete pathologic regression and others showing resistance to castration. As in humans, ADT decreased cell proliferation and prostate-specific antigen expression in TSGs. Adverse pathological features of parent HRPCa were associated with lack of regression of cancer in corresponding TSGs after ADT. Castration-resistant cancer cells remaining in TSGs showed upregulated expression of androgen receptor target genes, as occurs in castration-resistant prostate cancer (CRPC) in humans. Finally, a rare subset of castration-resistant cancer cells in TSGs underwent epithelial-mesenchymal transition, a process also observed in CRPC in humans. Conclusions: Our study demonstrates the feasibility of generating TSGs from multiple patients and of generating a relatively large number of TSGs from the same HRPCa specimen with similar cell composition and histology among control and experimental samples in an in vivo setting. The authentic response of TSGs to ADT, which has been extensively characterized in humans, suggests that TSGs can serve as a surrogate model for clinical trials to achieve rapid and less expensive screening of synthetic lethality based therapeutics for HRPCa and primary CRPC. Citation Format: Hongjuan Zhao, Alan Thong, Rosie Nolley, Stephen Reese, Jennifer Santos, Alexandre Ingels, Donna Peehl. Patient-derived Tissue Slice grafts of high-risk primary prostate cancer: An authentic preclinical model for synthetic lethality-based therapy. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Synthetic Lethal Approaches to Cancer Vulnerabilities; May 17-20, 2013; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(5 Suppl):Abstract nr B32.
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Tissue Slice grafts an in vivo model of human prostate androgen signaling
American Journal of Pathology, 2010Co-Authors: Hongjuan Zhao, Rosalie Nolley, Zuxiong Chen, Donna M. PeehlAbstract:We developed a Tissue Slice graft (TSG) model by implanting thin, precision-cut Tissue Slices derived from fresh primary prostatic adenocarcinomas under the renal capsule of immunodeficient mice. This new in vivo model not only allows analysis of approximately all of the cell types present in prostate cancer within an intact Tissue microenvironment, but also provides a more accurate assessment of the effects of interventions when Tissues from the same specimen with similar cell composition and histology are used as control and experimental samples. The thinness of the Slices ensures that sufficient samples can be obtained for large experiments as well as permits optimal exchange of nutrients, oxygen, and drugs between the grafted Tissue and the host. Both benign and cancer Tissues displayed characteristic histology and expression of cell-type specific markers for up to 3 months. Moreover, androgen-regulated protein expression diminished in TSGs after androgen ablation of the host and was restored after androgen repletion. Finally, many normal secretory epithelial cells and cancer cells in TSGs remained viable 2 months after androgen ablation, consistent with similar observations in postprostatectomy specimens following neoadjuvant androgen ablation. Among these were putative Nkx3.1+ stem cells. Our novel TSG model has the appropriate characteristics to serve as a useful tool to model all stages of disease, including normal Tissue, premalignant lesions, well-differentiated cancer, and poorly differentiated cancer.
Mariam A Stoffkhalili - One of the best experts on this subject based on the ideXlab platform.
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employment of liver Tissue Slice analysis to assay hepatotoxicity linked to replicative and nonreplicative adenoviral agents
Cancer Gene Therapy, 2006Co-Authors: Mariam A Stoffkhalili, Angel A Rivera, Alexander Stoff, Marieke Everts, Juan L Contreras, Dongquan Chen, L Teng, Marianne G Rots, Hidde J Haisma, Rodney P RocconiAbstract:Whereas virotherapy has emerged as a novel and promising approach for neoplastic diseases, appropriate model systems have hampered preclinical evaluation of candidate conditionally replicative adenovirus agents (CRAds) with respect to liver toxicity. This is due to the inability of human viral agents to cross species. We have recently shown the human liver Tissue Slice model to be a facile means to validate adenoviral replication. On this basis, we sought to determine whether our ex vivo liver Tissue Slice model could be used to assess CRAd-mediated liver toxicity. We analyzed and compared the toxicity of a conditionally replicative adenovirus (AdDelta24) to that of a replication incompetent adenovirus (Adnull [E1-]) in mouse and human liver Tissue Slices. To accomplish this, we examined the hepatic apoptosis expression profile by DNA microarray analyses, and compared these results to extracellular release of aminotransferase enzymes, along with direct evidence of apoptosis by caspase-3 immunhistochemical staining and TUNEL assays. Human and mouse liver Tissue Slices demonstrated a marked increase in extracellular release of aminotransferase enzymes on infection with AdDelta24 compared to Adnull. AdDelta24-mediated liver toxicity was further demonstrated by apoptosis induction, as detected by caspase-3 immunohistochemical staining, TUNEL assay and microarray analysis. In conclusion, concordance of CRAd-mediated apoptosis in both the human and the mouse liver Tissue Slice models was demonstrated, despite the limited replication ability of CRAds in mouse liver Slices. The results of this study, defining the CRAd-mediated apoptosis gene expression profiles in human and mouse liver, may lay a foundation for preclinical liver toxicity analysis of CRAd agents.
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a human adenoviral vector with a chimeric fiber from canine adenovirus type 1 results in novel expanded tropism for cancer gene therapy
Gene Therapy, 2005Co-Authors: Mariam A Stoffkhalili, Angel A Rivera, Joel N Glasgow, Alexander Stoff, Maaike Everts, Yuko Tsuruta, Yosuke Kawakami, Gerd J Bauerschmitz, James Michael Mathis, Larisa PereboevaAbstract:The development of novel therapeutic strategies is imperative for the treatment of advanced cancers like ovarian cancer and glioma, which are resistant to most traditional treatment modalities. In this regard, adenoviral (Ad) cancer gene therapy is a promising approach. However, the gene delivery efficiency of human serotype 5 recombinant adenoviruses (Ad5) in cancer gene therapy clinical trials to date has been limited, mainly due to the paucity of the primary Ad5 receptor, the coxsackie and adenovirus receptor (CAR), on human cancer cells. To circumvent CAR deficiency, Ad5 vectors have been retargeted by creating chimeric fibers possessing the knob domains of alternate human Ad serotypes. Recently, more radical modifications based on 'xenotype' knob switching with non-human adenovirus have been exploited. Herein, we present the characterization of a novel vector derived from a recombinant Ad5 vector containing the canine adenovirus serotype 1 (CAV-1) knob (Ad5Luc1-CK1), the tropism of which has not been previously described. We compared the function of this vector with our other chimeric viruses displaying the CAV-2 knob (Ad5Luc1-CK2) and Ad3 knob (Ad5/3Luc1). Our data demonstrate that the CAV-1 knob can alter Ad5 tropism through the use of a CAR-independent entry pathway distinct from that of both Ad5Luc1-CK2 and Ad5/3-Luc1. In fact, the gene transfer efficiency of this novel vector in ovarian cancer cell lines, and more importantly in patient ovarian cancer primary Tissue Slice samples, was superior relative to all other vectors applied in this study. Thus, CAV-1 knob xenotype gene transfer represents a viable means to achieve enhanced transduction of low-CAR tumors.
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preclinical evaluation of transcriptional targeting strategies for carcinoma of the breast in a Tissue Slice model system
Breast Cancer Research, 2005Co-Authors: Mariam A Stoffkhalili, Angel A Rivera, Alexander Stoff, Maaike Everts, Minghui Wang, Nilam Sanjib Banerjee, Scott Young, Gene P Siegal, Dirk F Richter, P DallAbstract:In view of the limited success of available treatment modalities for metastatic breast cancer, alternative and complementary strategies need to be developed. Adenoviral vector mediated strategies for breast cancer gene therapy and virotherapy are a promising novel therapeutic platform for the treatment of breast cancer. However, the promiscuous tropism of adenoviruses (Ads) is a major concern. Employing Tissue specific promoters (TSPs) to restrict transgene expression or viral replication is an effective way to increase specificity towards tumor Tissues and to reduce adverse effects in non-target Tissues such as the liver. In this regard, candidate breast cancer TSPs include promoters of the genes for the epithelial glycoprotein 2 (EGP-2), cyclooxygenase-2 (Cox-2), α-chemokine SDF-1 receptor (stromal-cell-derived factor, CXCR4), secretory leukoprotease inhibitor (SLPI) and survivin. We employed E1-deleted Ads that express the reporter gene luciferase under the control of the promoters of interest. We evaluated this class of vectors in various established breast cancer cell lines, primary breast cancer cells and finally in the most stringent preclinical available substrate system, constituted by precision cut Tissue Slices of human breast cancer and liver. Overall, the CXCR4 promoter exhibited the highest luciferase activity in breast cancer cell lines, primary breast cancer cells and breast cancer Tissue Slices. Importantly, the CXCR4 promoter displayed a very low activity in human primary fibroblasts and human liver Tissue Slices. Interestingly, gene expression profiles correlated with the promoter activities both in breast cancer cell lines and primary breast cancer cells. These data suggest that the CXCR4 promoter has an ideal 'breast cancer-on/liver-off' profile, and could, therefore, be a powerful tool in Ad vector based gene therapy or virotherapy of the carcinoma of the breast.
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gene transfer to carcinoma of the breast with fiber modified adenoviral vectors in a Tissue Slice model system
Cancer Biology & Therapy, 2005Co-Authors: Mariam A Stoffkhalili, Angel A Rivera, Alexander Stoff, Maaike Everts, Yosuke Kawakami, Michael J Mathis, Minghui Wang, Reinhard Waehler, Quiana L Mathews, Masato YamamotoAbstract:Successful adenoviral (Ad) vector-mediated strategies for breast cancer gene therapy and virotherapy have heretofore been hindered by low transduction efficiency. This has recently been understood to result from a relative paucity of expression of the primary adenovirus receptor, coxsackie-adenovirus-receptor (CAR), on primary tumor cells. To further investigate this issue, we evaluated the expression of CAR on breast cancer cell lines as well as primary breast cancer cells. With the exception of one patient sample, CAR expression was notably higher in the tumor cells from patients compared to CAR expression in the tumor cell lines. Furthermore, we explored CAR-independent targeting strategies to breast cancer Tissue by exploring a panel of infectivity-enhanced Ad vectors, which contain CAR-independent targeting motifs for their utility in breast cancer gene therapy and virotherapy. These targeting motifs included Ad 3 knob (Ad5/3), canine Ad serotype 2 knob (Ad5CAV-2), RGD (Ad5.RGD), polylysine (Ad5.pK7), or both RGD and polylysine (Ad5.RGD.pK7), and were tested using the breast cancer Tissue Slice model, which is the most stringent substrate system available. Of all the tested tropism modified Ad vectors, Ad5/3 exhibited the highest transductional efficiency in breast cancer. These preclinical results suggest that Ad5/3 is the most useful modification to achieve higher clinical efficacy of breast cancer gene therapy and virotherapy.
