| 1. |
- Kehoe, Laura, et al.
(author)
-
Make EU trade with Brazil sustainable
- 2019
-
In: Science. - : American Association for the Advancement of Science (AAAS). - 0036-8075 .- 1095-9203. ; 364:6438, s. 341-
-
Journal article (other academic/artistic)
|
|
| 2. |
|
|
| 3. |
- Agudo, Marta, et al.
(author)
-
Time course profiling of the retinal transcriptome after optic nerve transection and optic nerve crush
- 2008
-
In: Molecular Vision. - 1090-0535. ; 14, s. 1050-1063
-
Journal article (peer-reviewed)abstract
- PURPOSE:A time-course analysis of gene regulation in the adult rat retina after intraorbital nerve crush (IONC) and intraorbital nerve transection (IONT).METHODS:RNA was extracted from adult rat retinas undergoing either IONT or IONC at increasing times post-lesion. Affymetrix RAE230.2 arrays were hybridized and analyzed. Statistically regulated genes were annotated and functionally clustered. Arrays were validated by means of quantative reverse transcription polymerase chain reaction (qRT-PCR) on ten regulated genes at two times post-lesion. Western blotting and immunohistofluorescence for four pro-apoptotic proteins were performed on naïve and injured retinas. Finally, custom signaling maps for IONT- and IONC-induced death response were generated (MetaCore, Genego Inc.).RESULTS:Here we show that over time, 3,219 sequences were regulated after IONT and 1,996 after IONC. Out of the total of regulated sequences, 1,078 were commonly regulated by both injuries. Interestingly, while IONT mainly triggers a gene upregulation-sustained over time, IONC causes a transitory downregulation. Functional clustering identified the regulation of high interest biologic processes, most importantly cell death wherein apoptosis was the most significant cluster. Ten death-related genes upregulated by both injuries were used for array validation by means of qRT-PCR. In addition, western blotting and immunohistofluorescence of total and active Caspase 3 (Casp3), tumor necrosis factor receptor type 1 associated death domain (TRADD), tumor necrosis factor receptor superfamily member 1a (TNFR1a), and c-fos were performed to confirm their protein regulation and expression pattern in naïve and injured retinas. These analyses demonstrated that for these genes, protein regulation followed transcriptional regulation and that these pro-apoptotic proteins were expressed by retinal ganglion cells (RGCs). MetaCore-based death-signaling maps show that several apoptotic cascades were regulated in the retina following optic nerve injury and highlight the similarities and differences between IONT and IONC in cell death profiling.CONCLUSIONS: This comprehensive time course retinal transcriptome study comparing IONT and IONC lesions provides a unique valuable tool to understand the molecular mechanisms underlying optic nerve injury and to design neuroprotective protocols.
|
|
| 4. |
- Marin Navarro, Ana
(author)
-
Developing new models of childhood malignancies using human induced pluripotent stem cells
- 2019
-
Doctoral thesis (other academic/artistic)abstract
- Early onset diseases such as childhood malignancies and neurodevelopmental disorders have been intricate to study. For many years, research has relied and dependent upon different animal systems. Despite the usefulness of these systems, which have allowed the understanding of the biology behind these processes, the differences between species are still an undoubted fact. The emergence of induced pluripotent stem (iPS) cell technology has indeed opened new venues for many fields including disease modeling, personalized cell therapy, and drug screening. iPS cells have the potential to virtually differentiate into any cell type, hence becoming an unlimited source of disease-relevant cell types. Here, we present examples demonstrating the potential of disease modeling using patient-derived iPS cells. Neuroblastoma (NB) and Medulloblastoma (MB) are both cancers linked to dysregulations in pathways important during human development. Whereas NB develops during the peripheral nervous system (PNS) development, MB initiates during central nervous system (CNS) development. We have taken advantage of the early developmental signature of iPS cells to model cancer. We used non-cancerous cells from patients carrying germline mutations in cancer predisposing genes, ALK and PTCH1, and developed in vivo models that offer a unique understanding of cancer initiation and progression. NB patients carrying an ALK germline mutation were used to generate iPS cells (Paper I) and subsequent differentiation to Neural Crest Cells (NCC) was performed (Paper II, III). For this, a NCC generation protocol was optimized using intermediate levels of BMP (Paper II). Next, labelled NCC from patients and controls were orthotopically transplantated into the adrenal gland of immunodeficient mice (Paper III). Mice were followed in vivo using IVIS system, and we detected increased luciferase signal after more than 8 weeks but no signal was observed in mice injected with control NCC. After a year, adrenal glands from mice were harvested and one case of ganglioneuroblastoma was diagnosed, suggesting a low penetrance and mild phenotype of ALK contribution in NB initiation. Using a similar workflow, we generated Neuroepithelial stem (NES) cells from iPS cells derived from Gorlin syndrome patients. Gorlin patients carry germline mutations in PTCH1. Mutations in PTCH1 constitutively activate the Sonic Hedgehog (SHH) signalling pathway. In vivo transplantation of patient cells into the cerebellum of immunocompromised mice showed faithful resemblance of human SHH MB. By establishing NES cell cultures derived from MB tumors in the mice cerebellum, we could show the potential use of this model for identifying new targets for cancer treatment (Paper IV).b Moreover, we exploited 2D and 3D human in vitro systems derived from iPS cells to study the role of p53 during early brain development. We show that p53 has an important function in maintaining the appropriate structure of human brain organoids. Moreover, we demonstrate that p53 maintains genomic instability and primes neural differentiation in human NES cells. Thus, revealing the role of p53 in a human in vitro context of brain development (Paper V). In summary our work presents the big potential of iPS cell technology in the field of modeling disease.
|
|
| 5. |
- Susanto, Evelyn, et al.
(author)
-
Modeling SHH-driven medulloblastoma with patient iPS cell-derived neural stem cells
- 2020
-
In: Proceedings of the National Academy of Sciences of the United States of America. - : NATL ACAD SCIENCES. - 0027-8424 .- 1091-6490. ; 117:33, s. 20127-20138
-
Journal article (peer-reviewed)abstract
- Medulloblastoma is the most common malignant brain tumor in children. Here we describe a medulloblastoma model using In-duced pluripotent stem (iPS) cell-derived human neuroepithelial stem (NES) cells generated from a Gorlin syndrome patient carry-ing a germline mutation in the sonic hedgehog (SHH) receptor PTCH1. We found that Gorlin NES cells formed tumors in mouse cerebellum mimicking human medulloblastoma. Retransplantation of tumor-isolated NES (tNES) cells resulted in accelerated tumor formation, cells with reduced growth factor dependency, en-hanced neurosphere formation in vitro, and increased sensitivity to Vismodegib. Using our model, we identified LGALS1 to be a GLI target gene that is up-regulated in both Gorlin tNES cells and SHH-subgroup of medulloblastoma patients. Taken together, we dem-onstrate that NES cells derived from Gorlin patients can be used as a resource to model medulloblastoma initiation and progression and to identify putative targets.
|
|
