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Panaliappan, Tamilarasan K.
Alternative names
Publications (5 of 5) Show all publications
Panaliappan, T. K., Slekiene, L. & Gunhaga, L. (2022). CAM-Delam Assay to Score Metastatic Properties by Quantifying Delamination and Invasion Capacity of Cancer Cells. Journal of Visualized Experiments, 2022(184), Article ID e64025.
Open this publication in new window or tab >>CAM-Delam Assay to Score Metastatic Properties by Quantifying Delamination and Invasion Capacity of Cancer Cells
2022 (English)In: Journal of Visualized Experiments, E-ISSN 1940-087X, Vol. 2022, no 184, article id e64025Article in journal (Refereed) Published
Abstract [en]

The major cause of cancer-related deaths is metastasis formation (i.e., when cancer cells spread from the primary tumor to distant organs and form secondary tumors). Delamination, defined as the degradation of the basal lamina and basement membrane, is the initial process that facilitates the transmigration and spread of cancer cells to other tissues and organs. Scoring the delamination capacity of cancer cells would indicate the metastatic potential of these cells. We have developed a standardized method, the ex ovo CAM-Delam assay, to visualize and quantify the ability of cancer cells to delaminate and invade, thereby being able to assess metastatic aggressiveness. Briefly, the CAM-Delam method includes seeding cancer cells in silicone rings on the chick chorioallantoic membrane (CAM) at embryonic day 10, followed by incubation from hours to a few days. The CAM-Delam assay includes the use of an internal humidified chamber during chick embryo incubation. This novel approach increased embryo survival from 10%-50% to 80%-90%, which resolved previous technical problems with low embryo survival rates in different CAM assays. Next, the CAM samples with associated cancer cell clusters were isolated, fixed, and frozen. Finally, cryostat-sectioned samples were visualized and analyzed for basement membrane damage and cancer cell invasion using immunohistochemistry. By evaluating various known metastatic and non-metastatic cancer cell lines designed to express green fluorescent protein (GFP), the CAM-Delam quantitative results showed that the delamination capacity patterns reflect metastatic aggressiveness and can be scored into four categories. Future use of this assay, apart from quantifying delamination capacity as an indication of metastatic aggressiveness, is to unravel the molecular mechanisms that control delamination, invasion, the formation of micrometastases, and changes in the tumor microenvironment.

Place, publisher, year, edition, pages
MyJove Corporation, 2022
National Category
Cell and Molecular Biology Cancer and Oncology
Identifiers
urn:nbn:se:umu:diva-203195 (URN)10.3791/64025 (DOI)000898056900081 ()35723486 (PubMedID)2-s2.0-85132454671 (Scopus ID)
Funder
Swedish Cancer Society, 18 0463Swedish Research Council, 2017-01430
Available from: 2023-01-18 Created: 2023-01-18 Last updated: 2024-01-17
Wang, T., Sarwar, M., Whitchurch, J. B., Collins, H. M., Green, T., Semenas, J., . . . Persson, J. L. (2022). PIP5K1α is Required for Promoting Tumor Progression in Castration-Resistant Prostate Cancer. Frontiers in Cell and Developmental Biology, 10, Article ID 798590.
Open this publication in new window or tab >>PIP5K1α is Required for Promoting Tumor Progression in Castration-Resistant Prostate Cancer
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2022 (English)In: Frontiers in Cell and Developmental Biology, E-ISSN 2296-634X, Vol. 10, article id 798590Article in journal (Refereed) Published
Abstract [en]

PIP5K1α has emerged as a promising drug target for the treatment of castration-resistant prostate cancer (CRPC), as it acts upstream of the PI3K/AKT signaling pathway to promote prostate cancer (PCa) growth, survival and invasion. However, little is known of the molecular actions of PIP5K1α in this process. Here, we show that siRNA-mediated knockdown of PIP5K1α and blockade of PIP5K1α action using its small molecule inhibitor ISA-2011B suppress growth and invasion of CRPC cells. We demonstrate that targeted deletion of the N-terminal domain of PIP5K1α in CRPC cells results in reduced growth and migratory ability of cancer cells. Further, the xenograft tumors lacking the N-terminal domain of PIP5K1α exhibited reduced tumor growth and aggressiveness in xenograft mice as compared to that of controls. The N-terminal domain of PIP5K1α is required for regulation of mRNA expression and protein stability of PIP5K1α. This suggests that the expression and oncogenic activity of PIP5K1α are in part dependent on its N-terminal domain. We further show that PIP5K1α acts as an upstream regulator of the androgen receptor (AR) and AR target genes including CDK1 and MMP9 that are key factors promoting growth, survival and invasion of PCa cells. ISA-2011B exhibited a significant inhibitory effect on AR target genes including CDK1 and MMP9 in CRPC cells with wild-type PIP5K1α and in CRPC cells lacking the N-terminal domain of PIP5K1α. These results indicate that the growth of PIP5K1α-dependent tumors is in part dependent on the integrity of the N-terminal sequence of this kinase. Our study identifies a novel functional mechanism involving PIP5K1α, confirming that PIP5K1α is an intriguing target for cancer treatment, especially for treatment of CRPC.

Place, publisher, year, edition, pages
Frontiers Media S.A., 2022
Keywords
androgen receptor (AR), castration-resistant prostate cancer (CRPC), cyclin-dependent kinase (CDK), matrix metalloproteinases 9 (MMP9) PIP5K1α, phosphatidylinositol 4-phosphate 5 kinase (PIP5K1α), targeted therapy
National Category
Cancer and Oncology
Research subject
Oncology
Identifiers
urn:nbn:se:umu:diva-193614 (URN)10.3389/fcell.2022.798590 (DOI)000780059400001 ()35386201 (PubMedID)2-s2.0-85128078251 (Scopus ID)
Funder
EU, Horizon 2020, 721297Swedish Childhood Cancer Foundation, TJ2015-0097Swedish Cancer Society, CAN-2017-381Swedish Research Council, 2019-01318The Swedish Foundation for International Cooperation in Research and Higher Education (STINT), IG2013-5595The Kempe FoundationsCancerforskningsfonden i Norrland
Available from: 2022-04-07 Created: 2022-04-07 Last updated: 2023-05-23Bibliographically approved
Panaliappan, T. K., Slekiene, L., Jonasson, A.-K., Gilthorpe, J. D. & Gunhaga, L. (2020). CAM-Delam: an in vivo approach to visualize and quantify the delamination and invasion capacity of human cancer cells. Scientific Reports, 10(1), Article ID 10472.
Open this publication in new window or tab >>CAM-Delam: an in vivo approach to visualize and quantify the delamination and invasion capacity of human cancer cells
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2020 (English)In: Scientific Reports, E-ISSN 2045-2322, Vol. 10, no 1, article id 10472Article in journal (Refereed) Published
Abstract [en]

The development of metastases is the major cause of cancer related death. To develop a standardized method that define the ability of human cancer cells to degrade the basement membrane, e.g. the delamination capacity, is of importance to assess metastatic aggressiveness. We now present the in vivo CAM-Delam assay to visualize and quantify the ability of human cancer cells to delaminate and invade. The method includes seeding cancer cells on the chick chorioallantoic membrane (CAM), followed by the evaluation of cancer-induced delamination and potential invasion within hours to a few days. By testing a range of human cancer cell lines in the CAM-Delam assay, our results show that the delamination capacity can be divided into four categories and used to quantify metastatic aggressiveness. Our results emphasize the usefulness of this assay for quantifying delamination capacity as a measurement of metastatic aggressiveness, and in unraveling the molecular mechanisms that regulate delamination, invasion, formation of micro-metastases and modulations of the tumor microenvironment. This method will be useful in both the preclinical and clinical characterization of tumor biopsies, and in the validation of compounds that may improve survival in metastatic cancer.

Place, publisher, year, edition, pages
Nature Publishing Group, 2020
National Category
Cancer and Oncology
Research subject
Biological Research on Drug Dependence; Oncology
Identifiers
urn:nbn:se:umu:diva-173655 (URN)10.1038/s41598-020-67492-7 (DOI)000545967200069 ()32591581 (PubMedID)2-s2.0-85086871538 (Scopus ID)
Funder
Swedish Cancer Society, A549Swedish Cancer Society, SW620Swedish Cancer Society, 143BSwedish Cancer Society, 18 0463Cancerforskningsfonden i NorrlandSwedish Research Council, 2017-01430Knut and Alice Wallenberg Foundation, KAW2012.0091
Available from: 2020-07-21 Created: 2020-07-21 Last updated: 2024-04-10Bibliographically approved
Panaliappan, T. K., Slekiene, L., Gunhaga, L. & Patthey, C. (2019). Extensive apoptosis during the formation of the terminal nerve ganglion by olfactory placode-derived cells with distinct molecular markers. Differentiation, 110, 8-16
Open this publication in new window or tab >>Extensive apoptosis during the formation of the terminal nerve ganglion by olfactory placode-derived cells with distinct molecular markers
2019 (English)In: Differentiation, ISSN 0301-4681, E-ISSN 1432-0436, Vol. 110, p. 8-16Article in journal (Refereed) Published
Abstract [en]

The terminal nerve ganglion (TNG) is a well-known structure of the peripheral nervous system in cartilaginous and teleost fishes. It derives from the olfactory placode during embryonic development. While the differentiation and migration of gonadotropin releasing hormone (GnRH)-expressing neurons from the olfactory placode has been well documented, the TNG has been neglected in birds and mammals, and its development is less well described. Here we describe the formation of a ganglion-like structure from migratory olfactory placodal cells in chicken. The TNG is surrounded by neural crest cells, but in contrast to other cranial sensory ganglia, we observed no neural crest corridor, and olfactory unsheathing cells appear only after the onset of neuronal migration. We identified Isl1 and Lhx2 as two transcription factors that label neuronal subpopulations in the forming TNG, distinct from GnRH1(+) cells, thereby revealing a diversity of cell types during the formation of the TNG. We also provide evidence for extensive apoptosis in the terminal nerve ganglion shortly after its formation, but not in other cranial sensory ganglia. Moreover, at later stages placode-derived neurons expressing GnRH1, Isl1 and/or Lhx2 become incorporated in the telencephalon. The integration of TNG neurons into the telencephalon together with the earlier widespread apoptosis in the TNG might be an explanation why the TNG in mammals and birds is much smaller compared to other vertebrates.

Place, publisher, year, edition, pages
Elsevier, 2019
Keywords
Terminal nerve ganglion, Apoptosis, Lhx2, GnRH, Isl1, Chicken
National Category
Neurosciences
Identifiers
urn:nbn:se:umu:diva-167041 (URN)10.1016/j.diff.2019.09.003 (DOI)000498845100002 ()31539705 (PubMedID)2-s2.0-85072197946 (Scopus ID)
Funder
Swedish Research Council, 2017-01430_3
Available from: 2020-01-08 Created: 2020-01-08 Last updated: 2023-03-24Bibliographically approved
Panaliappan, T. K., Wittmann, W., Jidigam, V. K., Mercurio, S., Bertolini, J. A., Sghari, S., . . . Gunhaga, L. (2018). Sox2 is required for olfactory pit formation and olfactory neurogenesis through BMP restriction and Hes5 upregulation. Development, 145(2), Article ID dev153791.
Open this publication in new window or tab >>Sox2 is required for olfactory pit formation and olfactory neurogenesis through BMP restriction and Hes5 upregulation
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2018 (English)In: Development, ISSN 0950-1991, E-ISSN 1477-9129, Vol. 145, no 2, article id dev153791Article in journal (Refereed) Published
Abstract [en]

The transcription factor Sox2 is necessary to maintain pluripotency of embryonic stem cells, and to regulate neural development. Neurogenesis in the vertebrate olfactory epithelium persists from embryonic stages through adulthood. The role Sox2 plays for the development of the olfactory epithelium and neurogenesis within has, however, not been determined. Here, by analysing Sox2 conditional knockout mouse embryos and chick embryos deprived of Sox2 in the olfactory epithelium using CRISPR-Cas9, we show that Sox2 activity is crucial for the induction of the neural progenitor gene Hes5 and for subsequent differentiation of the neuronal lineage. Our results also suggest that Sox2 activity promotes the neurogenic domain in the nasal epithelium by restricting Bmp4 expression. The Sox2-deficient olfactory epithelium displays diminished cell cycle progression and proliferation, a dramatic increase in apoptosis and finally olfactory pit atrophy. Moreover, chromatin immunoprecipitation data show that Sox2 directly binds to the Hes5 promoter in both the PNS and CNS. Taken together, our results indicate that Sox2 is essential to establish, maintain and expand the neuronal progenitor pool by suppressing Bmp4 and upregulating Hes5 expression.

Place, publisher, year, edition, pages
Company of Biologists Ltd, 2018
Keywords
Sox2, Hes5, Olfactory epithelium, Neurogenesis, Mouse
National Category
Developmental Biology Cell and Molecular Biology
Identifiers
urn:nbn:se:umu:diva-145166 (URN)10.1242/dev.153791 (DOI)000424412000009 ()2-s2.0-85041301091 (Scopus ID)
Available from: 2018-02-23 Created: 2018-02-23 Last updated: 2023-03-24Bibliographically approved
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