{"id":{"repo_id":"kings","oai_identifier":"oai:kclpure.kcl.ac.uk:studenttheses/8d963ed0-7b21-4f73-bc94-ec5a76205bdd"},"canonical_url":"https://search.dev.ndltd.org/etd/kings/oai:kclpure.kcl.ac.uk:studenttheses/8d963ed0-7b21-4f73-bc94-ec5a76205bdd","repository":{"repo_id":"kings","name":"King's College London","base_url":"https://kclpure.kcl.ac.uk/ws/oai"},"display":{"title":"T-type calcium channels and human mesangial cell proliferation","abstract":"Aberrant proliferation of human mesangial cells (MC) is a critical step in the pathogenesis of mesangioproliferative renal diseases. The T-type calcium channel (T-CaCN) has been proposed to play an important role in the proliferation of a number of non-excitable cell types, but T-CaCN expression and functional significance in MC is not known. The aim of this thesis was to investigate the hypothesis that T-CaCN may play an important role in the proliferation of MC in primary culture.
Expression of mRNA encoding the α1H isoform (Cav3.2 clone) (but not the α1G nor α1I T-CaCN isoforms) was demonstrated in human MC by RT-PCR. Expression of α1H T-CaCN protein was difficult to assess directly due to the lack of a highly sensitive and specific antibody, despite attempts at upregulation of the protein. Electrophysiological studies of MC demonstrated an inward calcium current in a proportion of cells with characteristics consistent with T-CaCN. Culture of MC with the T-CaCN antagonists mibefradil, Ni2+ and TTL-1177 resulted in a significant reduction in the growth velocity of MC. This effect was not seen upon incubation with verapamil, an L-type calcium channel antagonist. DNA synthesis in MC treated with each of the T-CaCN antagonists was significantly reduced by up to 50% as shown by BrdU incorporation. This anti-proliferative effect was not associated with direct drug-induced cytotoxicity or apoptosis. FACS analysis of MC incubated with T-CaCN antagonists illustrated an increased proportion of cells remaining in G1 and not progressing into S-phase. Treatment of cultured MC with TTL-1177 resulted in a significant reduction in the signalling protein p-ERK within 30 minutes, an effect not seen with verapamil, suggesting a possible mechanism needing further investigation. MC transfection with siRNA targeting the α1H isoform resulted in significant knockdown of T-CaCN α1H mRNA and a reduction in the growth velocity of cultured MC of approximately 50% compared to control siRNA transfection, with an associated 37% reduction in DNA synthesis.
These results demonstrate evidence for an important role for T-type calcium channels in the proliferation of human mesangial cells, justifying further study in in greater detail. This could potentially lead to a novel therapy in the treatment of proliferative renal diseases.","abstract_html":"Aberrant proliferation of human mesangial cells (MC) is a critical step in the pathogenesis of mesangioproliferative renal diseases. The T-type calcium channel (T-CaCN) has been proposed to play an important role in the proliferation of a number of non-excitable cell types, but T-CaCN expression and functional significance in MC is not known. The aim of this thesis was to investigate the hypothesis that T-CaCN may play an important role in the proliferation of MC in primary culture. <br/><br/>Expression of mRNA encoding the α1H isoform (Cav3.2 clone) (but not the α1G nor α1I T-CaCN isoforms) was demonstrated in human MC by RT-PCR. Expression of α1H T-CaCN protein was difficult to assess directly due to the lack of a highly sensitive and specific antibody, despite attempts at upregulation of the protein. Electrophysiological studies of MC demonstrated an inward calcium current in a proportion of cells with characteristics consistent with T-CaCN. Culture of MC with the T-CaCN antagonists mibefradil, Ni2+ and TTL-1177 resulted in a significant reduction in the growth velocity of MC. This effect was not seen upon incubation with verapamil, an L-type calcium channel antagonist. DNA synthesis in MC treated with each of the T-CaCN antagonists was significantly reduced by up to 50% as shown by BrdU incorporation. This anti-proliferative effect was not associated with direct drug-induced cytotoxicity or apoptosis. FACS analysis of MC incubated with T-CaCN antagonists illustrated an increased proportion of cells remaining in G1 and not progressing into S-phase. Treatment of cultured MC with TTL-1177 resulted in a significant reduction in the signalling protein p-ERK within 30 minutes, an effect not seen with verapamil, suggesting a possible mechanism needing further investigation. MC transfection with siRNA targeting the α1H isoform resulted in significant knockdown of T-CaCN α1H mRNA and a reduction in the growth velocity of cultured MC of approximately 50% compared to control siRNA transfection, with an associated 37% reduction in DNA synthesis.<br/><br/>These results demonstrate evidence for an important role for T-type calcium channels in the proliferation of human mesangial cells, justifying further study in in greater detail. This could potentially lead to a novel therapy in the treatment of proliferative renal diseases.","abstract_has_math":false,"creators":["Mulgrew, Christopher James"],"institution":"King's College London","degree_name":"Doctor of Philosophy","degree_level":"Doctoral Thesis","degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":["Hendry, Bruce Melville","Shattock, Michael Jonathan"],"committee_chairs":[],"committee_members":[],"year":2012,"date_issued":"2012-6-1","date_published":"2012-6-1","updated_at":"2026-07-24T02:44:40Z","subjects":[],"languages":["eng"],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["oai:kclpure.kcl.ac.uk:studenttheses/8d963ed0-7b21-4f73-bc94-ec5a76205bdd"],"render_values":[{"text":"oai:kclpure.kcl.ac.uk:studenttheses/8d963ed0-7b21-4f73-bc94-ec5a76205bdd","href":null,"code":true}]}]},"links":{"outbound_url":"https://kclpure.kcl.ac.uk/portal/en/studentTheses/8d963ed0-7b21-4f73-bc94-ec5a76205bdd","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Hendry, Bruce Melville","Shattock, Michael Jonathan"]},{"key":"dc:creator","label":"Author","values":["Mulgrew, Christopher James"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2012-6-1"]},{"key":"dc:date.issued","label":"Date","values":["2012-6-1"]},{"key":"dc:publisher.department","label":"Dc Publisher Department","values":["Renal Sciences"]},{"key":"dc:publisher.institution","label":"Dc Publisher Institution","values":["King's College London"]},{"key":"dc:relation.isreferencedby","label":"Dc Relation Isreferencedby","values":["https://kclpure.kcl.ac.uk/portal/en/studentTheses/8d963ed0-7b21-4f73-bc94-ec5a76205bdd"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]},{"key":"dc:type.qualificationlevel","label":"Dc Type Qualificationlevel","values":["Doctoral Thesis"]},{"key":"dc:type.qualificationname","label":"Dc Type Qualificationname","values":["Doctor of Philosophy"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["oai:kclpure.kcl.ac.uk:studenttheses/8d963ed0-7b21-4f73-bc94-ec5a76205bdd","https://kclpure.kcl.ac.uk/portal/en/studentTheses/8d963ed0-7b21-4f73-bc94-ec5a76205bdd"]},{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://kclpure.kcl.ac.uk/portal/files/13136798/Studentthesis-Christopher%20James_Mulgrew_2012.pdf"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Aberrant proliferation of human mesangial cells (MC) is a critical step in the pathogenesis of mesangioproliferative renal diseases. The T-type calcium channel (T-CaCN) has been proposed to play an important role in the proliferation of a number of non-excitable cell types, but T-CaCN expression and functional significance in MC is not known. The aim of this thesis was to investigate the hypothesis that T-CaCN may play an important role in the proliferation of MC in primary culture.
Expression of mRNA encoding the α1H isoform (Cav3.2 clone) (but not the α1G nor α1I T-CaCN isoforms) was demonstrated in human MC by RT-PCR. Expression of α1H T-CaCN protein was difficult to assess directly due to the lack of a highly sensitive and specific antibody, despite attempts at upregulation of the protein. Electrophysiological studies of MC demonstrated an inward calcium current in a proportion of cells with characteristics consistent with T-CaCN. Culture of MC with the T-CaCN antagonists mibefradil, Ni2+ and TTL-1177 resulted in a significant reduction in the growth velocity of MC. This effect was not seen upon incubation with verapamil, an L-type calcium channel antagonist. DNA synthesis in MC treated with each of the T-CaCN antagonists was significantly reduced by up to 50% as shown by BrdU incorporation. This anti-proliferative effect was not associated with direct drug-induced cytotoxicity or apoptosis. FACS analysis of MC incubated with T-CaCN antagonists illustrated an increased proportion of cells remaining in G1 and not progressing into S-phase. Treatment of cultured MC with TTL-1177 resulted in a significant reduction in the signalling protein p-ERK within 30 minutes, an effect not seen with verapamil, suggesting a possible mechanism needing further investigation. MC transfection with siRNA targeting the α1H isoform resulted in significant knockdown of T-CaCN α1H mRNA and a reduction in the growth velocity of cultured MC of approximately 50% compared to control siRNA transfection, with an associated 37% reduction in DNA synthesis.
These results demonstrate evidence for an important role for T-type calcium channels in the proliferation of human mesangial cells, justifying further study in in greater detail. This could potentially lead to a novel therapy in the treatment of proliferative renal diseases."]},{"key":"dc:title","label":"Title","values":["T-type calcium channels and human mesangial cell proliferation"]}]}],"canonical_facts":{"dc:contributor.advisor":["Hendry, Bruce Melville","Shattock, Michael Jonathan"],"dc:creator":["Mulgrew, Christopher James"],"dc:date":["2012-6-1"],"dc:date.issued":["2012-6-1"],"dc:description.abstract":["Aberrant proliferation of human mesangial cells (MC) is a critical step in the pathogenesis of mesangioproliferative renal diseases. The T-type calcium channel (T-CaCN) has been proposed to play an important role in the proliferation of a number of non-excitable cell types, but T-CaCN expression and functional significance in MC is not known. The aim of this thesis was to investigate the hypothesis that T-CaCN may play an important role in the proliferation of MC in primary culture.
Expression of mRNA encoding the α1H isoform (Cav3.2 clone) (but not the α1G nor α1I T-CaCN isoforms) was demonstrated in human MC by RT-PCR. Expression of α1H T-CaCN protein was difficult to assess directly due to the lack of a highly sensitive and specific antibody, despite attempts at upregulation of the protein. Electrophysiological studies of MC demonstrated an inward calcium current in a proportion of cells with characteristics consistent with T-CaCN. Culture of MC with the T-CaCN antagonists mibefradil, Ni2+ and TTL-1177 resulted in a significant reduction in the growth velocity of MC. This effect was not seen upon incubation with verapamil, an L-type calcium channel antagonist. DNA synthesis in MC treated with each of the T-CaCN antagonists was significantly reduced by up to 50% as shown by BrdU incorporation. This anti-proliferative effect was not associated with direct drug-induced cytotoxicity or apoptosis. FACS analysis of MC incubated with T-CaCN antagonists illustrated an increased proportion of cells remaining in G1 and not progressing into S-phase. Treatment of cultured MC with TTL-1177 resulted in a significant reduction in the signalling protein p-ERK within 30 minutes, an effect not seen with verapamil, suggesting a possible mechanism needing further investigation. MC transfection with siRNA targeting the α1H isoform resulted in significant knockdown of T-CaCN α1H mRNA and a reduction in the growth velocity of cultured MC of approximately 50% compared to control siRNA transfection, with an associated 37% reduction in DNA synthesis.
These results demonstrate evidence for an important role for T-type calcium channels in the proliferation of human mesangial cells, justifying further study in in greater detail. This could potentially lead to a novel therapy in the treatment of proliferative renal diseases."],"dc:identifier":["oai:kclpure.kcl.ac.uk:studenttheses/8d963ed0-7b21-4f73-bc94-ec5a76205bdd","https://kclpure.kcl.ac.uk/portal/en/studentTheses/8d963ed0-7b21-4f73-bc94-ec5a76205bdd"],"dc:identifier.uri":["https://kclpure.kcl.ac.uk/portal/files/13136798/Studentthesis-Christopher%20James_Mulgrew_2012.pdf"],"dc:language":["eng"],"dc:publisher.department":["Renal Sciences"],"dc:publisher.institution":["King's College London"],"dc:relation.isreferencedby":["https://kclpure.kcl.ac.uk/portal/en/studentTheses/8d963ed0-7b21-4f73-bc94-ec5a76205bdd"],"dc:title":["T-type calcium channels and human mesangial cell proliferation"],"dc:type":["Thesis"],"dc:type.qualificationlevel":["Doctoral Thesis"],"dc:type.qualificationname":["Doctor of Philosophy"]},"updated_at":"2026-07-24T02:44:40Z"}