225 results on '"Donnelly, PS"'
Search Results
2. Novel Anti-Neuroinflammatory Properties of a Thiosemicarbazone-Pyridylhydrazone Copper(II) Complex
- Author
-
Choo, XY, McInnes, LE, Grubman, A, Wasielewska, JM, Belaya, I, Burrows, E, Quek, H, Martin, JC, Loppi, S, Sorvari, A, Rait, D, Powell, A, Duncan, C, Liddell, JR, Tanila, H, Polo, JM, Malm, T, Kanninen, KM, Donnelly, PS, White, AR, Choo, XY, McInnes, LE, Grubman, A, Wasielewska, JM, Belaya, I, Burrows, E, Quek, H, Martin, JC, Loppi, S, Sorvari, A, Rait, D, Powell, A, Duncan, C, Liddell, JR, Tanila, H, Polo, JM, Malm, T, Kanninen, KM, Donnelly, PS, and White, AR
- Abstract
Neuroinflammation has a major role in several brain disorders including Alzheimer's disease (AD), yet at present there are no effective anti-neuroinflammatory therapeutics available. Copper(II) complexes of bis(thiosemicarbazones) (CuII(gtsm) and CuII(atsm)) have broad therapeutic actions in preclinical models of neurodegeneration, with CuII(atsm) demonstrating beneficial outcomes on neuroinflammatory markers in vitro and in vivo. These findings suggest that copper(II) complexes could be harnessed as a new approach to modulate immune function in neurodegenerative diseases. In this study, we examined the anti-neuroinflammatory action of several low-molecular-weight, charge-neutral and lipophilic copper(II) complexes. Our analysis revealed that one compound, a thiosemicarbazone-pyridylhydrazone copper(II) complex (CuL5), delivered copper into cells in vitro and increased the concentration of copper in the brain in vivo. In a primary murine microglia culture, CuL5 was shown to decrease secretion of pro-inflammatory cytokine macrophage chemoattractant protein 1 (MCP-1) and expression of tumor necrosis factor alpha (Tnf), increase expression of metallothionein (Mt1), and modulate expression of Alzheimer's disease-associated risk genes, Trem2 and Cd33. CuL5 also improved the phagocytic function of microglia in vitro. In 5xFAD model AD mice, treatment with CuL5 led to an improved performance in a spatial working memory test, while, interestingly, increased accumulation of amyloid plaques in treated mice. These findings demonstrate that CuL5 can induce anti-neuroinflammatory effects in vitro and provide selective benefit in vivo. The outcomes provide further support for the development of copper-based compounds to modulate neuroinflammation in brain diseases.
- Published
- 2022
3. Hexadentate technetium-99m bis(thiosemicarbazonato) complexes: synthesis, characterisation and biodistribution
- Author
-
Kelderman, CAA, Davey, PRWJ, Ma, MT, de Veer, M, Salimova, E, Donnelly, PS, Paterson, BM, Kelderman, CAA, Davey, PRWJ, Ma, MT, de Veer, M, Salimova, E, Donnelly, PS, and Paterson, BM
- Abstract
The syntheses of non-oxido/non-nitrido bis(thiosemicarbazonato)technetium(V) complexes featuring a series of alkyl and ether substituents is presented. The bis(thiosemicarbazones) were radiolabelled with technetium-99m using an optimised one-pot synthesis from [99mTc][TcO4]-. Mass spectrometry and computational chemistry data suggested a distorted trigonal prismatic coordination environment for the bis(thiosemicarbazonato)technetium(V) complexes by way of a bis(thiosemicarbazone)technetium(V)-oxido intermediate complex. The lipophilicities of the complexes were estimated using distribution ratios and three of the new complexes were investigated in mice using kinetic planar imaging and ex vivo biodistribution experiments and were compared to [99mTc][TcO4]-. Modification of the technetium complexes with various lipophilic functional groups altered the biodistributions of the complexes in mice despite evidence suggesting limited stability of the complexes to biologically relevant conditions. The most hydrophilic complex had higher uptake in the kidneys compared to the most lipophilic, which had higher liver uptake, suggesting modification of the excretion pathways.
- Published
- 2022
4. Palladium-Mediated CO2 Extrusion Followed by Insertion of Allenes: Translating Mechanistic Studies to Develop a One-Pot Method for the Synthesis of Alkenes
- Author
-
Yang, Y, Spyrou, B, White, JM, Canty, AJ, Donnelly, PS, O'Hair, RAJ, Yang, Y, Spyrou, B, White, JM, Canty, AJ, Donnelly, PS, and O'Hair, RAJ
- Published
- 2022
5. Synthesis of acyloin natural products by Mukaiyama hydration
- Author
-
Ricca, M, Zhang, W, Li, J, Fellowes, T, White, JM, Donnelly, PS, Rizzacasa, MA, Ricca, M, Zhang, W, Li, J, Fellowes, T, White, JM, Donnelly, PS, and Rizzacasa, MA
- Abstract
The acyloin natural products are a family of bioactive compounds isolated from fungi and myxobacteria. The total synthesis of 7 members of the acyloin family was achieved via a HWE reaction followed by Mukaiyama-Isayama hydration, using novel Co(II) and Co(III) Schiff base SALPN complexes as catalysts for the key enone hydration step. Furthermore, we have shown that a mild acyloin rearrangement is possible under Mukaiyama hydration conditions, which was crucial in the success of this approach.
- Published
- 2022
6. ImmunoPET: IMaging of cancer imMUNOtherapy targets with positron Emission Tomography: a phase 0/1 study characterising PD-L1 with 89Zr-durvalumab (MEDI4736) PET/CT in stage III NSCLC patients receiving chemoradiation study protocol.
- Author
-
Hegi-Johnson, F, Rudd, SE, Wichmann, C, Akhurst, T, Roselt, P, Trinh, J, John, T, Devereux, L, Donnelly, PS, Hicks, R, Scott, AM, Steinfort, D, Fox, S, Blyth, B, Parakh, S, Hanna, GG, Callahan, J, Burbury, K, MacManus, M, Hegi-Johnson, F, Rudd, SE, Wichmann, C, Akhurst, T, Roselt, P, Trinh, J, John, T, Devereux, L, Donnelly, PS, Hicks, R, Scott, AM, Steinfort, D, Fox, S, Blyth, B, Parakh, S, Hanna, GG, Callahan, J, Burbury, K, and MacManus, M
- Abstract
BACKGROUND: ImmunoPET is a multicentre, single arm, phase 0-1 study that aims to establish if 89Zr-durvalumab PET/CT can be used to interrogate the expression of PD-L1 in larger, multicentre clinical trials. METHODS: The phase 0 study recruited 5 PD-L1+ patients with metastatic non-small cell lung cancer (NSCLC). Patients received 60MBq/70 kg 89Zr-durva up to a maximum of 74 MBq, with scan acquisition at days 0, 1, 3 or 5±1 day. Data on (1) Percentage of injected 89Zr-durva dose found in organs of interest (2) Absorbed organ doses (µSv/MBq of administered 89Zr-durva) and (3) whole-body dose expressed as mSv/100MBq of administered dose was collected to characterise biodistribution.The phase 1 study will recruit 20 patients undergoing concurrent chemoradiotherapy for stage III NSCLC. Patients will have 89Zr-durva and FDG-PET/CT before, during and after chemoradiation. In order to establish the feasibility of 89Zr-durva PET/CT for larger multicentre trials, we will collect both imaging and toxicity data. Feasibility will be deemed to have been met if more than 80% of patients are able complete all trial requirements with no significant toxicity. ETHICS AND DISSEMINATION: This phase 0 study has ethics approval (HREC/65450/PMCC 20/100) and is registered on the Australian Clinical Trials Network (ACTRN12621000171819). The protocol, technical and clinical data will be disseminated by conference presentations and publications. Any modifications to the protocol will be formally documented by administrative letters and must be submitted to the approving HREC for review and approval. TRIAL REGISTRATION NUMBER: Australian Clinical Trials Network ACTRN12621000171819.
- Published
- 2022
7. Positron Emission Tomographic Imaging of Tumor Cell Death Using Zirconium-89-Labeled APOMAB (R) Following Cisplatin Chemotherapy in Lung and Ovarian Cancer Xenograft Models
- Author
-
Liapis, V, Tieu, W, Wittwer, NL, Gargett, T, Evdokiou, A, Takhar, P, Rudd, SE, Donnelly, PS, Brown, MP, Staudacher, AH, Liapis, V, Tieu, W, Wittwer, NL, Gargett, T, Evdokiou, A, Takhar, P, Rudd, SE, Donnelly, PS, Brown, MP, and Staudacher, AH
- Abstract
PURPOSE: Early detection of tumor treatment responses represents an unmet clinical need with no approved noninvasive methods. DAB4, or its chimeric derivative, chDAB4 (APOMAB®) is an antibody that targets the Lupus associated antigen (La/SSB). La/SSB is over-expressed in malignancy and selectively targeted by chDAB4 in cancer cells dying from DNA-damaging treatment. Therefore, chDAB4 is a unique diagnostic tool that detects dead cancer cells and thus could distinguish between treatment responsive and nonresponsive patients. PROCEDURES: In clinically relevant tumor models, mice bearing subcutaneous xenografts of human ovarian or lung cancer cell lines or intraperitoneal ovarian cancer xenografts were untreated or given chemotherapy followed 24h later by chDAB4 radiolabeled with [89Zr]ZrIV. Tumor responses were monitored using bioluminescence imaging and caliper measurements. [89Zr]Zr-chDAB4 uptake in tumor and normal tissues was measured using an Albira SI Positron-Emission Tomography (PET) imager and its biodistribution was measured using a Hidex gamma-counter. RESULTS: Tumor uptake of [89Zr]Zr-chDAB4 was detected in untreated mice, and uptake significantly increased in both human lung and ovarian tumors after chemotherapy, but not in normal tissues. CONCLUSION: Given that tumors, rather than normal tissues, were targeted after chemotherapy, these results support the clinical development of chDAB4 as a radiodiagnostic imaging agent and as a potential predictive marker of treatment response.
- Published
- 2021
8. A clinical trial of non-invasive imaging with an anti-HIV antibody labelled with copper-64 in people living with HIV and uninfected controls
- Author
-
McMahon, JH, Zerbato, JM, Lau, JSY, Lange, JL, Roche, M, Tumpach, C, Dantanarayana, A, Rhodes, A, Chang, J, Rasmussen, TA, Mackenzie, CA, Alt, K, Hagenauer, M, Roney, J, O'Bryan, J, Carey, A, McIntyre, R, Beech, P, O'Keefe, GJ, Wichmann, CW, Scott, FE, Guo, N, Lee, S-T, Liu, Z, Caskey, M, Nussenzweig, MC, Donnelly, PS, Egan, G, Hagemeyer, CE, Scott, AM, Lewin, SR, McMahon, JH, Zerbato, JM, Lau, JSY, Lange, JL, Roche, M, Tumpach, C, Dantanarayana, A, Rhodes, A, Chang, J, Rasmussen, TA, Mackenzie, CA, Alt, K, Hagenauer, M, Roney, J, O'Bryan, J, Carey, A, McIntyre, R, Beech, P, O'Keefe, GJ, Wichmann, CW, Scott, FE, Guo, N, Lee, S-T, Liu, Z, Caskey, M, Nussenzweig, MC, Donnelly, PS, Egan, G, Hagemeyer, CE, Scott, AM, and Lewin, SR
- Abstract
BACKGROUND: A research priority in finding a cure for HIV is to establish methods to accurately locate and quantify where and how HIV persists in people living with HIV (PLWH) receiving suppressive antiretroviral therapy (ART). Infusing copper-64 (64Cu) radiolabelled broadly neutralising antibodies targeting HIV envelope (Env) with CT scan and positron emission tomography (PET) identified HIV Env in tissues in SIV infected non-human primates . We aimed to determine if a similar approach was effective in people living with HIV (PLWH). METHODS: Unmodified 3BNC117 was compared with 3BNC117 bound to the chelator MeCOSar and 64Cu (64Cu-3BNC117) in vitro to assess binding and neutralization. In a clinical trial 64Cu-3BNC117 was infused into HIV uninfected (Group 1), HIV infected and viremic (viral load, VL >1000 c/mL; Group 2) and HIV infected aviremic (VL <20 c/mL; Group 3) participants using two dosing strategies: high protein (3mg/kg unlabeled 3BNC117 combined with <5mg 64Cu-3BNC117) and trace (<5mg 64Cu-3BNC117 only). All participants were screened for 3BNC117 sensitivity from virus obtained from viral outgrowth. Magnetic resonance imaging (MRI)/PET and pharmacokinetic assessments (ELISA for serum 3BNC117 concentrations and gamma counting for 64Cu) were performed 1, 24- and 48-hours post dosing. The trial (clincialtrials.gov NCT03063788) primary endpoint was comparison of PET standard uptake values (SUVs) in regions of interest (e.g lymph node groups and gastrointestinal tract). FINDINGS: Comparison of unmodified and modified 3BNC117 in vitro demonstrated no difference in HIV binding or neutralisation. 17 individuals were enrolled of which 12 were dosed including Group 1 (n=4, 2 high protein, 2 trace dose), Group 2 (n=6, 2 high protein, 4 trace) and Group 3 (n=2, trace only). HIV+ participants had a mean CD4 of 574 cells/microL and mean age 43 years. There were no drug related adverse effects and no differences in tissue uptake in regions of interest (e.g lymph node g
- Published
- 2021
9. CuATSM improves motor function and extends survival but is not tolerated at a high dose in SOD1(G93A) mice with a C57BL/6 background
- Author
-
Lum, JS, Brown, ML, Farrawell, NE, McAlary, L, Ly, D, Chisholm, CG, Snow, J, Vine, KL, Karl, T, Kreilaus, F, McInnes, LE, Nikseresht, S, Donnelly, PS, Crouch, PJ, Yerbury, JJ, Lum, JS, Brown, ML, Farrawell, NE, McAlary, L, Ly, D, Chisholm, CG, Snow, J, Vine, KL, Karl, T, Kreilaus, F, McInnes, LE, Nikseresht, S, Donnelly, PS, Crouch, PJ, and Yerbury, JJ
- Abstract
The synthetic copper-containing compound, CuATSM, has emerged as one of the most promising drug candidates developed for the treatment of amyotrophic lateral sclerosis (ALS). Multiple studies have reported CuATSM treatment provides therapeutic efficacy in various mouse models of ALS without any observable adverse effects. Moreover, recent results from an open label clinical study suggested that daily oral dosing with CuATSM slows disease progression in patients with both sporadic and familial ALS, providing encouraging support for CuATSM in the treatment of ALS. Here, we assessed CuATSM in high copy SOD1G93A mice on the congenic C57BL/6 background, treating at 100 mg/kg/day by gavage, starting at 70 days of age. This dose in this specific model has not been assessed previously. Unexpectedly, we report a subset of mice initially administered CuATSM exhibited signs of clinical toxicity, that necessitated euthanasia in extremis after 3-51 days of treatment. Following a 1-week washout period, the remaining mice resumed treatment at the reduced dose of 60 mg/kg/day. At this revised dose, treatment with CuATSM slowed disease progression and increased survival relative to vehicle-treated littermates. This work provides the first evidence that CuATSM produces positive disease-modifying outcomes in high copy SOD1G93A mice on a congenic C57BL/6 background. Furthermore, results from the 100 mg/kg/day phase of the study support dose escalation determination of tolerability as a prudent step when assessing treatments in previously unassessed models or genetic backgrounds.
- Published
- 2021
10. Enzyme mediated incorporation of zirconium-89 or copper-64 into a fragment antibody for same day imaging of epidermal growth factor receptor
- Author
-
Rudd, SE, Van Zuylekom, JK, Raicevic, A, Pearce, LA, Cullinane, C, Williams, CC, Adams, TE, Hicks, RJ, Donnelly, PS, Rudd, SE, Van Zuylekom, JK, Raicevic, A, Pearce, LA, Cullinane, C, Williams, CC, Adams, TE, Hicks, RJ, and Donnelly, PS
- Abstract
Identification of tumors which over-express Epidermal Growth Factor Receptor (EGFR) is important in selecting patients for anti-EGFR therapies. Enzymatic bioconjugation was used to introduce positron-emitting radionuclides (89Zr, 64Cu) into an anti-EGFR antibody fragment for Positron Emission Tomography (PET) imaging the same day as injection. A monovalent antibody fragment with high affinity for EGFR was engineered to include a sequence that is recognized by the transpeptidase sortase A. Two different metal chelators, one for 89ZrIV and one for 64CuII, were modified with a N-terminal glycine to enable them to act as substrates in sortase A mediated bioconjugation to the antibody fragment. Both fragments provided high-quality PET images of EGFR positive tumors in a mouse model at 3 hours post-injection, a significant advantage when compared to radiolabeled full antibodies that require several days between injection of the tracer and imaging. The use of enzymatic bioconjugation gives reproducible homogeneous products with the metal complexes selectively installed on the C-terminus of the antibody potentially simplifying regulatory approval.
- Published
- 2021
11. Harnessing Cu-64/Cu-67 for a theranostic approach to pretargeted radioimmunotherapy
- Author
-
Keinanen, O, Fung, K, Brennan, JM, Zia, N, Harris, M, van Dam, E, Biggin, C, Hedt, A, Stoner, J, Donnelly, PS, Lewis, JS, Zeglis, BM, Keinanen, O, Fung, K, Brennan, JM, Zia, N, Harris, M, van Dam, E, Biggin, C, Hedt, A, Stoner, J, Donnelly, PS, Lewis, JS, and Zeglis, BM
- Abstract
Over the past decade, theranostic imaging has emerged as a powerful clinical tool in oncology for identifying patients likely to respond to targeted therapies and for monitoring the response of patients to treatment. Herein, we report a theranostic approach to pretargeted radioimmunotherapy (PRIT) based on a pair of radioisotopes of copper: positron-emitting copper-64 (64Cu, t1/2 = 12.7 h) and beta particle-emitting copper-67 (67Cu, t1/2 = 61.8 h). This strategy is predicated on the in vivo ligation between a trans-cyclooctene (TCO)-bearing antibody and a tetrazine (Tz)-based radioligand via the rapid and bioorthogonal inverse electron-demand Diels–Alder reaction. Longitudinal therapy studies were conducted in a murine model of human colorectal carcinoma using an immunoconjugate of the huA33 antibody modified with TCO (huA33-TCO) and a 67Cu-labeled Tz radioligand ([67Cu]Cu-MeCOSar-Tz). The injection of huA33-TCO followed 72 h later by the administration of 18.5, 37.0, or 55.5 MBq of [67Cu]Cu-MeCOSar-Tz produced a dose-dependent therapeutic response, with the median survival time increasing from 68 d for the lowest dose to >200 d for the highest. Furthermore, we observed that mice that received the highest dose of [67Cu]Cu-MeCOSar-Tz in a fractionated manner exhibited improved hematological values without sacrificing therapeutic efficacy. Dual radionuclide experiments in which a single administration of huA33-TCO was followed by separate injections of [64Cu]Cu-MeCOSar-Tz and [67Cu]Cu-MeCOSar-Tz revealed that the positron emission tomography images produced by the former accurately predicted the efficacy of the latter. In these experiments, a correlation was observed between the tumoral uptake of [64Cu]Cu-MeCOSar-Tz and the subsequent therapeutic response to [67Cu]Cu-MeCOSar-Tz.
- Published
- 2020
12. Improved non-invasive positron emission tomographic imaging of chemotherapy-induced tumor cell death using Zirconium-89-labeled APOMAB (R)
- Author
-
Liapis, V, Tieu, W, Rudd, SE, Donnelly, PS, Wittwer, NL, Brown, MP, Staudacher, AH, Liapis, V, Tieu, W, Rudd, SE, Donnelly, PS, Wittwer, NL, Brown, MP, and Staudacher, AH
- Abstract
PURPOSE: The chimeric monoclonal antibody (mAb) chDAB4 (APOMAB®) targets the Lupus associated (La)/Sjögren Syndrome-B (SSB) antigen, which is over-expressed in tumors but only becomes available for antibody binding in dead tumor cells. Hence, chDAB4 may be used as a novel theranostic tool to distinguish between responders and nonresponders early after chemotherapy. Here, we aimed to ascertain which positron emitter, Zirconium-89 ([89Zr]ZrIV) or Iodine-124 ([124I]I), was best suited to label chDAB4 for post-chemotherapy PET imaging of tumor-bearing mice and to determine which of two different bifunctional chelators provided optimal tumor imaging by PET using [89Zr]ZrIV-labeled chDAB4. METHODS: C57BL/6 J mice bearing subcutaneous syngeneic tumors of EL4 lymphoma were either untreated or given chemotherapy, then administered radiolabeled chDAB4 after 24 h with its biodistribution examined using PET and organ assay. We compared chDAB4 radiolabeled with [89Zr] ZrIV or [124I] I, or [89Zr]Zr-chDAB4 using either DFO-NCS or DFOSq as a chelator. RESULTS: After chemotherapy, [89Zr]Zr-chDAB4 showed higher and prolonged mean (± SD) tumor uptake of 29.5 ± 5.9 compared to 7.8 ± 1.2 for [124I] I -chDAB4. In contrast, antibody uptake in healthy tissues was not affected. Compared to DFO-NCS, DFOSq did not result in significant differences in tumor uptake of [89Zr]Zr-chDAB4 but did alter the tumor:liver ratio in treated mice 3 days after injection in favour of DFOSq (8.0 ± 1.1) compared to DFO-NCS (4.2 ± 0.7). CONCLUSION: ImmunoPET using chDAB4 radiolabeled with residualizing [89Zr] ZrIV rather than [124I] I optimized post-chemotherapy tumor uptake. Further, PET imaging characteristics were improved by DFOSq rather than DFO-NCS. Therefore, the radionuclide/chelator combination of [89Zr] ZrIV and DFOSq is preferred for the imminent clinical evaluation of chDAB4 as a selective tumor cell death radioligand.
- Published
- 2020
13. Cu-II(atsm) inhibits ferroptosis: Implications for treatment of neurodegenerative disease
- Author
-
Southon, A, Szostak, K, Acevedo, KM, Dent, KA, Volitakis, I, Belaidi, AA, Barnham, KJ, Crouch, PJ, Ayton, S, Donnelly, PS, Bush, A, Southon, A, Szostak, K, Acevedo, KM, Dent, KA, Volitakis, I, Belaidi, AA, Barnham, KJ, Crouch, PJ, Ayton, S, Donnelly, PS, and Bush, A
- Abstract
BACKGROUND AND PURPOSE: Diacetyl-bis(4-methyl-3-thiosemicarbazonato)copperII (CuII (atsm)) ameliorates neurodegeneration and delays disease progression in mouse models of amyotrophic lateral sclerosis (ALS) and Parkinson's disease (PD), yet the mechanism of action remains uncertain. Promising results were recently reported for separate Phase 1 studies in ALS patients and PD patients. Affected tissue in these disorders shares features of elevated Fe, low glutathione and increased lipid peroxidation consistent with ferroptosis, a novel form of regulated cell death. We therefore evaluated the ability of CuII (atsm) to inhibit ferroptosis. EXPERIMENTAL APPROACH: Ferroptosis was induced in neuronal cell models by inhibition of glutathione peroxidase-4 activity with RSL3 or by blocking cystine uptake with erastin. Cell viability and lipid peroxidation were assessed and the efficacy of CuII (atsm) was compared to the known antiferroptotic compound liproxstatin-1. KEY RESULTS: CuII (atsm) protected against lipid peroxidation and ferroptotic lethality in primary and immortalised neuronal cell models (EC50 : ≈130 nM, within an order of magnitude of liproxstatin-1). NiII (atsm) also prevented ferroptosis with similar potency, whereas ionic CuII did not. In cell-free systems, CuII (atsm) and NiII (atsm) inhibited FeII -induced lipid peroxidation, consistent with these compounds quenching lipid radicals. CONCLUSIONS AND IMPLICATIONS: The antiferroptotic activity of CuII (atsm) could therefore be the disease-modifying mechanism being tested in ALS and PD trials. With potency in vitro approaching that of liproxstatin-1, CuII (atsm) possesses favourable properties such as oral bioavailability and entry into the brain that make it an attractive investigational product for clinical trials of ferroptosis-related diseases.
- Published
- 2020
14. Gallium Fluoride Complexes with Acyclic Bispicolinic Ligands as Potential New Fluorine-18 Labelled Imaging Agents
- Author
-
Koay, H, Haskali, MB, Roselt, PD, White, JM, Donnelly, PS, Koay, H, Haskali, MB, Roselt, PD, White, JM, and Donnelly, PS
- Published
- 2020
15. Site-Specific Glycation and Chemo-enzymatic Antibody Sortagging for the Retargeting of rAAV6 to Inflamed Endothelium
- Author
-
Pearce, HA, Qian, H, Connell, Timothy, Huang, D, Gottstein, C, Donnelly, PS, Peter, K, Gregorevic, P, Hagemeyer, CE, Pearce, HA, Qian, H, Connell, Timothy, Huang, D, Gottstein, C, Donnelly, PS, Peter, K, Gregorevic, P, and Hagemeyer, CE
- Published
- 2019
16. A conceptual framework for the development of iridium(iii) complex-based electrogenerated chemiluminescence labels
- Author
-
Chen, L, Hayne, DJ, Doeven, EH, Agugiaro, J, Wilson, DJD, Henderson, LC, Connell, TU, Nai, YH, Alexander, R, Carrara, S, Hogan, CF, Donnelly, PS, Francis, PS, Chen, L, Hayne, DJ, Doeven, EH, Agugiaro, J, Wilson, DJD, Henderson, LC, Connell, TU, Nai, YH, Alexander, R, Carrara, S, Hogan, CF, Donnelly, PS, and Francis, PS
- Abstract
Translation of the highly promising electrogenerated chemiluminescence (ECL) properties of Ir(iii) complexes (with tri-n-propylamine (TPrA) as a co-reactant) into a new generation of ECL labels for ligand binding assays necessitates the introduction of functionality suitable for bioconjugation. Modification of the ligands, however, can affect not only the photophysical and electrochemical properties of the complex, but also the reaction pathways available to generate light. Through a combined theoretical and experimental study, we reveal the limitations of conventional approaches to the design of electrochemiluminophores and introduce a new class of ECL label, [Ir(C^N)2(pt-TOxT-Sq)]+ (where C^N is a range of possible cyclometalating ligands, and pt-TOxT-Sq is a pyridyltriazole ligand with trioxatridecane chain and squarate amide ethyl ester), which outperformed commercial Ir(iii) complex labels in two commonly used assay formats. Predicted limits on the redox potentials and emission wavelengths of Ir(iii) complexes capable of generating ECL via the dominant pathway applicable in microbead supported ECL assays were experimentally verified by measuring the ECL intensities of the parent luminophores at different applied potentials, and comparing the ECL responses for the corresponding labels under assay conditions. This study provides a framework to tailor ECL labels for specific assay conditions and a fundamental understanding of the ECL pathways that will underpin exploration of new luminophores and co-reactants.
- Published
- 2019
17. A Bivalent Inhibitor of Prostate Specific Membrane Antigen Radiolabeled with Copper‐64 with High Tumor Uptake and Retention
- Author
-
Zia, NA, Cullinane, C, Van Zuylekom, JK, Waldeck, K, McInnes, LE, Buncic, G, Haskali, MB, Roselt, PD, Hicks, RJ, Donnelly, PS, Zia, NA, Cullinane, C, Van Zuylekom, JK, Waldeck, K, McInnes, LE, Buncic, G, Haskali, MB, Roselt, PD, Hicks, RJ, and Donnelly, PS
- Published
- 2019
18. Modulating Targeting of Poly(ethylene glycol) Particles to Tumor Cells Using Bispecific Antibodies
- Author
-
Cui, J, Ju, Y, Houston, ZH, Class, JJ, Fletcher, NL, Alcantara, S, Dai, Q, Howard, CB, Mahler, SM, Wheatley, AK, De Rose, R, Brannon, PT, Paterson, BM, Donnelly, PS, Thurecht, K, Caruso, F, Kent, SJ, Cui, J, Ju, Y, Houston, ZH, Class, JJ, Fletcher, NL, Alcantara, S, Dai, Q, Howard, CB, Mahler, SM, Wheatley, AK, De Rose, R, Brannon, PT, Paterson, BM, Donnelly, PS, Thurecht, K, Caruso, F, and Kent, SJ
- Abstract
Low-fouling or "stealth" particles composed of poly(ethylene glycol) (PEG) display a striking ability to evade phagocytic cell uptake. However, functionalizing them for specific targeting is challenging. To address this challenge, stealth PEG particles prepared by a mesoporous silica templating method are functionalized with bispecific antibodies (BsAbs) to obtain PEG-BsAb particles via a one-step binding strategy for cell and tumor targeting. The dual specificity of the BsAbs-one arm binds to the PEG particles while the other targets a cell antigen (epidermal growth factor receptor, EGFR)-is exploited to modulate the number of targeting ligands per particle. Increasing the BsAb incubation concentration increases the amount of BsAb tethered to the PEG particles and enhances targeting and internalization into breast cancer cells overexpressing EGFR. The degree of BsAb functionalization does not significantly reduce the stealth properties of the PEG particles ex vivo, as assessed by their interactions with primary human blood granulocytes and monocytes. Although increasing the BsAb amount on PEG particles does not lead to the expected improvement in tumor accumulation in vivo, BsAb functionalization facilitates tumor cell uptake of PEG particles. This work highlights strategies to balance evading nonspecific clearance pathways, while improving tumor targeting and accumulation.
- Published
- 2019
19. The social defeat/overcrowding murine psychosocial stress model results in a pharmacologically reversible body weight gain but not depression - related behaviours
- Author
-
Keenan, RJ, Chan, J, Donnelly, PS, Barnham, KJ, Jacobson, LH, Keenan, RJ, Chan, J, Donnelly, PS, Barnham, KJ, and Jacobson, LH
- Abstract
Depression is a highly prevalent psychiatric disorder, yet its etiology is not well understood. The validation of animal models is therefore a critical step towards advancing knowledge about the neurobiology of depression. Psychosocial stress has been promoted as a prospective animal model of depression, however, different protocols exist with variable responses, and further investigations are therefore required. We aimed to characterise the behavioural and body weight responses to the social defeat/overcrowding (SD/OC) model and to explore the effects of the antidepressant fluoxetine and the peroxynitrite scavenger, CuII(atsm), therein. Male C57BL/6JArc mice were exposed to a 19 day SD/OC protocol at two levels of aggression, determined by terminating SD bouts after one, or approximately five social defeat postures. This was followed by a battery of behavioural tests including social interaction test (SIT), locomotor activity (LMA), light-dark box test (LDB), saccharin preference test (SPT) and the forced swim test (FST). Mice were dosed daily with vehicle, fluoxetine (20 mg/kg) or CuII(atsm) (30 mg/kg) throughout the protocol. SD/OC increased body weight compared to controls, which was abolished by fluoxetine and attenuated by CuII(atsm). Weight gain specifically peaked during OC sessions but was not affected by either drug treatment. Fluoxetine reduced the number of defeat postures during fight bouts on some days. SD/OC otherwise failed to elicit depression- or anxiety-like behaviour in the tests measured. These data raise questions over the SD/OC model as an etiological model of depression-related behaviours but highlight the potential of this model for investigations into mechanisms regulating binge eating and weight gain under conditions of chronic social stress.
- Published
- 2018
20. Cu-II(atsm) Attenuates Neuroinflammation
- Author
-
Choo, XY, Liddell, JR, Huuskonen, MT, Grubman, A, Moujalled, D, Roberts, J, Kysenius, K, Patten, L, Quek, H, Oikari, LE, Duncan, C, James, SA, McInnes, LE, Hayne, DJ, Donnelly, PS, Pollari, E, Vahatalo, S, Lejavova, K, Kettunen, M, Malm, T, Koistinaho, J, White, AR, Kanninen, KM, Choo, XY, Liddell, JR, Huuskonen, MT, Grubman, A, Moujalled, D, Roberts, J, Kysenius, K, Patten, L, Quek, H, Oikari, LE, Duncan, C, James, SA, McInnes, LE, Hayne, DJ, Donnelly, PS, Pollari, E, Vahatalo, S, Lejavova, K, Kettunen, M, Malm, T, Koistinaho, J, White, AR, and Kanninen, KM
- Abstract
Background: Neuroinflammation and biometal dyshomeostasis are key pathological features of several neurodegenerative diseases, including Alzheimer's disease (AD). Inflammation and biometals are linked at the molecular level through regulation of metal buffering proteins such as the metallothioneins. Even though the molecular connections between metals and inflammation have been demonstrated, little information exists on the effect of copper modulation on brain inflammation. Methods: We demonstrate the immunomodulatory potential of the copper bis(thiosemicarbazone) complex CuII(atsm) in an neuroinflammatory model in vivo and describe its anti-inflammatory effects on microglia and astrocytes in vitro. Results: By using a sophisticated in vivo magnetic resonance imaging (MRI) approach, we report the efficacy of CuII(atsm) in reducing acute cerebrovascular inflammation caused by peripheral administration of bacterial lipopolysaccharide (LPS). CuII(atsm) also induced anti-inflammatory outcomes in primary microglia [significant reductions in nitric oxide (NO), monocyte chemoattractant protein 1 (MCP-1), and tumor necrosis factor (TNF)] and astrocytes [significantly reduced NO, MCP-1, and interleukin 6 (IL-6)] in vitro. These anti-inflammatory actions were associated with increased cellular copper levels and increased the neuroprotective protein metallothionein-1 (MT1) in microglia and astrocytes. Conclusion: The beneficial effects of CuII(atsm) on the neuroimmune system suggest copper complexes are potential therapeutics for the treatment of neuroinflammatory conditions.
- Published
- 2018
21. The Copper bis(thiosemicarbazone) Complex Cu-II(atsm) Is Protective Against Cerebral Ischemia Through Modulation of the Inflammatory Milieu
- Author
-
Huuskonen, MT, Tuo, Q-Z, Loppi, S, Dhungana, H, Korhonen, P, McInnes, LE, Donnelly, PS, Grubman, A, Wojciechowski, S, Lejavova, K, Pomeshchik, Y, Periviita, L, Kosonen, L, Giordano, M, Walker, FR, Liu, R, Bush, AI, Koistinaho, J, Malm, T, White, AR, Lei, P, Kanninen, KM, Huuskonen, MT, Tuo, Q-Z, Loppi, S, Dhungana, H, Korhonen, P, McInnes, LE, Donnelly, PS, Grubman, A, Wojciechowski, S, Lejavova, K, Pomeshchik, Y, Periviita, L, Kosonen, L, Giordano, M, Walker, FR, Liu, R, Bush, AI, Koistinaho, J, Malm, T, White, AR, Lei, P, and Kanninen, KM
- Abstract
Developing new therapies for stroke is urgently needed, as this disease is the leading cause of death and disability worldwide, and the existing treatment is only available for a small subset of patients. The interruption of blood flow to the brain during ischemic stroke launches multiple immune responses, characterized by infiltration of peripheral immune cells, the activation of brain microglial cells, and the accumulation of immune mediators. Copper is an essential trace element that is required for many critical processes in the brain. Copper homeostasis is disturbed in chronic neurodegenerative diseases and altered in stroke patients, and targeted copper delivery has been shown to be protective against chronic neurodegeneration. This study was undertaken to assess whether the copper bis(thiosemicarbazone) complex, CuII(atsm), is beneficial in acute brain injury, in preclinical mouse models of ischemic stroke. We demonstrate that the copper complex CuII(atsm) protects neurons from excitotoxicity and N2a cells from OGD in vitro, and is protective in permanent and transient ischemia models in mice as measured by functional outcome and lesion size. Copper delivery in the ischemic brains modulates the inflammatory response, specifically affecting the myeloid cells. It reduces CD45 and Iba1 immunoreactivity, and alters the morphology of Iba1 positive cells in the ischemic brain. CuII(atsm) also protects endogenous microglia against ischemic insult and reduces the proportion of invading monocytes. These results demonstrate that the copper complex CuII(atsm) is an inflammation-modulating compound with high therapeutic potential in stroke and is a strong candidate for the development of therapies for acute brain injury.
- Published
- 2017
22. Guest‐induced Assembly of Bis(thiosemicarbazonato) Zinc(II) Coordination Nanotubes
- Author
-
Paterson, BM, White, KF, White, JM, Abrahams, BF, Donnelly, PS, Paterson, BM, White, KF, White, JM, Abrahams, BF, and Donnelly, PS
- Published
- 2017
23. Cu-II(atsm) improves the neurological phenotype and survival of SOD1(G93A) mice and selectively increases enzymatically active SOD1 in the spinal cord
- Author
-
Hilton, JB, Mercer, SW, Lim, NKH, Faux, NG, Buncic, G, Beckman, JS, Roberts, BR, Donnelly, PS, White, AR, Crouch, PJ, Hilton, JB, Mercer, SW, Lim, NKH, Faux, NG, Buncic, G, Beckman, JS, Roberts, BR, Donnelly, PS, White, AR, and Crouch, PJ
- Abstract
Ubiquitous expression of mutant Cu/Zn-superoxide dismutase (SOD1) selectively affects motor neurons in the central nervous system (CNS), causing the adult-onset degenerative disease amyotrophic lateral sclerosis (ALS). The CNS-specific impact of ubiquitous mutant SOD1 expression is recapitulated in transgenic mouse models of the disease. Here we present outcomes for the metallo-complex CuII(atsm) tested for therapeutic efficacy in mice expressing SOD1G93A on a mixed genetic background. Oral administration of CuII(atsm) delayed the onset of neurological symptoms, improved locomotive capacity and extended overall survival. Although the ALS-like phenotype of SOD1G93A mice is instigated by expression of the mutant SOD1, we show the improved phenotype of the CuII(atsm)-treated animals involves an increase in mature mutant SOD1 protein in the disease-affected spinal cord, where concomitant increases in copper and SOD1 activity are also evident. In contrast to these effects in the spinal cord, treating with CuII(atsm) had no effect in liver on either mutant SOD1 protein levels or its activity, indicating a CNS-selective SOD1 response to the drug. These data provide support for CuII(atsm) as a treatment option for ALS as well as insight to the CNS-selective effects of mutant SOD1.
- Published
- 2017
24. Electron paramagnetic resonance microscopy using spins in diamond under ambient conditions
- Author
-
Simpson, DA, Ryan, RG, Hall, LT, Panchenko, E, Drew, SC, Petrou, S, Donnelly, PS, Mulvaney, P, Hollenberg, LCL, Simpson, DA, Ryan, RG, Hall, LT, Panchenko, E, Drew, SC, Petrou, S, Donnelly, PS, Mulvaney, P, and Hollenberg, LCL
- Abstract
Magnetic resonance spectroscopy is one of the most important tools in chemical and bio-medical research. However, sensitivity limitations typically restrict imaging resolution to ~ 10 µm. Here we bring quantum control to the detection of chemical systems to demonstrate high-resolution electron spin imaging using the quantum properties of an array of nitrogen-vacancy centres in diamond. Our electron paramagnetic resonance microscope selectively images electronic spin species by precisely tuning a magnetic field to bring the quantum probes into resonance with the external target spins. This provides diffraction limited spatial resolution of the target spin species over a field of view of 50 × 50 µm2 with a spin sensitivity of 104 spins per voxel or ∼100 zmol. The ability to perform spectroscopy and dynamically monitor spin-dependent redox reactions at these scales enables the development of electron spin resonance and zepto-chemistry in the physical and life sciences.Electron paramagnetic resonance spectroscopy has important scientific and medical uses but improving the resolution of conventional methods requires cryogenic, vacuum environments. Simpson et al. show nitrogen vacancy centres can be used for sub-micronmetre imaging with improved sensitivity in ambient conditions.
- Published
- 2017
25. Targeting Activated Platelets: A Unique and Potentially Universal Approach for Cancer Imaging
- Author
-
Yap, ML, McFadyen, JD, Wang, X, Zia, NA, Hohmann, JD, Ziegler, M, Yao, Y, Pham, A, Harris, M, Donnelly, PS, Hogarth, PM, Pietersz, GA, Lim, B, Peter, K, Yap, ML, McFadyen, JD, Wang, X, Zia, NA, Hohmann, JD, Ziegler, M, Yao, Y, Pham, A, Harris, M, Donnelly, PS, Hogarth, PM, Pietersz, GA, Lim, B, and Peter, K
- Abstract
Rationale The early detection of primary tumours and metastatic disease is vital for successful therapy and is contingent upon highly specific molecular markers and sensitive, non-invasive imaging techniques. We hypothesized that the accumulation of activated platelets within tumours is a general phenomenon and thus represents a novel means for the molecular imaging of cancer. Here we investigate a unique single chain antibody (scFv), which specifically targets activated platelets, as a novel biotechnological tool for molecular imaging of cancer. Methods The scFvGPIIb/IIIa, which binds specifically to the activated form of the platelet integrin receptor GPIIb/IIIa present on activated platelets, was conjugated to either Cy7, 64Cu or ultrasound-enhancing microbubbles. Using the Cy7 labelled scFvGPIIb/IIIa, fluorescence imaging was performed in mice bearing four different human tumour xenograft models; SKBr3, MDA-MB-231, Ramos and HT-1080 cells. Molecular imaging via PET and ultrasound was performed using the scFvGPIIb/IIIa-64Cu and scFvGPIIb/IIIa-microbubbles, respectively, to further confirm specific targeting of scFvGPIIb/IIIa to activated platelets in the tumour stroma. Results Using scFvGPIIb/IIIa we successfully showed specific targeting of activated platelets within the microenvironment of human tumour xenografts models via three different molecular imaging modalities. The presence of platelets within the tumour microenvironment, and as such their relevance as a molecular target epitope in cancer was further confirmed via immunofluorescence of human tumour sections of various cancer types, thus validating the translational importance of our novel approach to human disease. Conclusion Our study provides proof of concept for imaging and localization of tumours by molecular targeting activated platelets. We illustrate the utility of a unique scFv as a versatile biotechnological tool which can be conjugated to various contrast agents for molecular imaging of cancer
- Published
- 2017
26. Tunable Porous Coordination Polymers for the Capture, Recovery and Storage of Inhalation Anesthetics
- Author
-
Abrahams, BF, Dharma, AD, Donnelly, PS, Hudson, TA, Kepert, CJ, Robson, R, Southon, PD, White, KF, Abrahams, BF, Dharma, AD, Donnelly, PS, Hudson, TA, Kepert, CJ, Robson, R, Southon, PD, and White, KF
- Abstract
The uptake of inhalation anesthetics by three topologically identical frameworks is described. The 3D network materials, which possess square channels of different dimensions, are formed from the relatively simple combination of ZnII centres and dianionic ligands that contain a phenolate and a carboxylate group at opposite ends. All three framework materials are able to adsorb N2O, Xe and isoflurane. Whereas the framework with the widest channels is able to adsorb large quantities of the various guests from the gas phase, the frameworks with the narrower channels have superior binding enthalpies and exhibit higher levels of retention. The use of ligands in which substituents are bound to the aromatic rings of the bridging ligands offers great scope for tuning the adsorption properties of the framework materials.
- Published
- 2017
27. New perspectives on the annihilation electrogenerated chemiluminescence of mixed metal complexes in solution
- Author
-
Kerr, E, Doeven, EH, Barbante, GJ, Hogan, CF, Hayne, DJ, Donnelly, PS, Francis, PS, Kerr, E, Doeven, EH, Barbante, GJ, Hogan, CF, Hayne, DJ, Donnelly, PS, and Francis, PS
- Abstract
Preliminary explorations of the annihilation electrogenerated chemiluminescence (ECL) of mixed metal complexes have revealed opportunities to enhance emission intensities and control the relative intensities from multiple luminophores through the applied potentials. However, the mechanisms of these systems are only poorly understood. Herein, we present a comprehensive characterisation of the annihilation ECL of mixtures of tris(2,2'-bipyridine)ruthenium(ii) hexafluorophosphate ([Ru(bpy)3](PF6)2) and fac-tris(2-phenylpyridine)iridium(iii) ([Ir(ppy)3]). This includes a detailed investigation of the change in emission intensity from each luminophore as a function of both the applied electrochemical potentials and the relative concentrations of the two complexes, and a direct comparison with two mixed (Ru/Ir) ECL systems for which emission from only the ruthenium-complex was previously reported. Concomitant emission from both luminophores was observed in all three systems, but only when: (1) the applied potentials were sufficient to generate the intermediates required to form the electronically excited state of both complexes; and (2) the concentration of the iridium complex (relative to the ruthenium complex) was sufficient to overcome quenching processes. Both enhancement and quenching of the ECL of the ruthenium complex was observed, depending on the experimental conditions. The observations were rationalised through several complementary mechanisms, including resonance energy transfer and various energetically favourable electron-transfer pathways.
- Published
- 2016
28. Synthesis of Homochiral Co-III- and Mn-IV-[2.2]Paracyclophane Schiff Base Complexes with Predetermined Chirality at the Metal Centre
- Author
-
Loits, D, Braese, S, North, AJ, White, JM, Donnelly, PS, Rizzacasa, MA, Loits, D, Braese, S, North, AJ, White, JM, Donnelly, PS, and Rizzacasa, MA
- Published
- 2016
29. Oral treatment with CuII(atsm) increases mutant SOD1 in vivo but protects motor neurons and improves the phenotype of a transgenic mouse model of amyotrophic lateral sclerosis
- Author
-
Roberts, BR, Lim, NKH, McAllum, EJ, Donnelly, PS, Hare, DJ, Doble, PA, Turner, BJ, Price, KA, Lim, SC, Paterson, BM, Hickey, JL, Rhoads, TW, Williams, JR, Kanninen, KM, Hung, LW, Liddell, JR, Grubman, A, Monty, JF, Llanos, RM, Kramer, DR, Mercer, JFB, Bush, AI, Masters, CL, Duce, JA, Li, QX, Beckman, JS, Barnham, KJ, White, AR, and Crouch, PJ
- Subjects
Motor Neurons ,Thiosemicarbazones ,Neurology & Neurosurgery ,Superoxide Dismutase ,animal diseases ,Amyotrophic Lateral Sclerosis ,Age Factors ,nutritional and metabolic diseases ,Administration, Oral ,Mice, Transgenic ,nervous system diseases ,Mice ,Disease Models, Animal ,Phenotype ,Superoxide Dismutase-1 ,nervous system ,Spinal Cord ,Mutation ,Organometallic Compounds ,Chromatography, Gel ,Animals ,Humans ,Cation Transport Proteins ,Locomotion ,Copper Transporter 1 - Abstract
Mutations in the metallo-protein Cu/Zn-superoxide dismutase (SOD1) cause amyotrophic lateral sclerosis (ALS) in humans and an expression level-dependent phenotype in transgenic rodents. We show that oral treatment with the therapeutic agent diacetyl-bis(4-methylthiosemicarbazonato)copperII [CuII(atsm)] increased the concentration of mutant SOD1 (SOD1G37R) in ALS model mice, but paradoxically improved locomotor function and survival of the mice. To determine why the mice with increased levels of mutant SOD1 had an improved phenotype, we analyzed tissues by mass spectrometry. These analyses revealed most SOD1 in the spinal cord tissue of the SOD1G37R mice was Cu deficient. Treating with CuII(atsm) decreased the pool of Cu-deficient SOD1 and increased the pool of fully metallated (holo) SOD1. Tracking isotopically enriched 65CuII(atsm) confirmed the increase in holo-SOD1 involved transfer of Cu from CuII(atsm) to SOD1, suggesting the improved locomotor function and survival of the CuII(atsm)-treated SOD1G37R mice involved, at least in part, the ability of the compound to improve the Cu content of the mutant SOD1. This was supported by improved survival of SOD1G37R mice that expressed the human gene for the Cu uptake protein CTR1. Improving the metal content of mutant SOD1 in vivo with CuII(atsm) did not decrease levels of misfolded SOD1. These outcomes indicate the metal content of SOD1 may be a greater determinant of the toxicity of the protein in mutant SOD1-associated forms of ALS than the mutations themselves. Improving the metal content of SOD1 therefore represents a valid therapeutic strategy for treating ALS caused by SOD1. © 2014 the authors.
- Published
- 2014
30. PBT2 inhibits glutamate-induced excitotoxicity in neurons through metal-mediated preconditioning
- Author
-
Johanssen, T, Suphantarida, N, Donnelly, PS, Liu, XM, Petrou, S, Hill, AF, Barnham, KJ, Johanssen, T, Suphantarida, N, Donnelly, PS, Liu, XM, Petrou, S, Hill, AF, and Barnham, KJ
- Abstract
Excitotoxicity is the pathological process by which neuronal death occurs as a result of excessive stimulation of receptors at the excitatory synapse such as the NMDA receptor (NMDAR). Excitotoxicity has been implicated in the acute neurological damage from ischemia and traumatic brain injury and in the chronic neurodegeneration in Alzheimer's disease (AD) and Huntington's disease (HD). As a result NMDAR antagonists have become an attractive therapeutic strategy for the potential treatment of multiple neurodegenerative diseases. However NMDAR signaling is dichotomous in nature, with excessive increases in neuronal intracellular calcium through excessive NMDAR activity being lethal but moderate increases to intracellular calcium levels during normal synaptic function providing neuroprotection. Subsequently indiscriminant inhibition of this receptor is best avoided as was concluded from previous clinical trials of NMDAR antagonists. We show that the metal chaperone, PBT2, currently in clinical trials for HD, is able to protect against glutamate-induced excitotoxicity mediated through NMDARs. This was achieved by PBT2 inducing Zn(2+)-dependent increases in intracellular Ca(2+) levels resulting in preconditioning of neurons and inhibition of Ca(2+)-induced neurotoxic signaling cascade involving calpain-activated cleavage of calcineurin. Our study demonstrates that modulating intracellular Ca(2+) levels by a zinc ionophore is a valid therapeutic strategy to protect against the effects of excitotoxicity thought to underlie both acute and chronic neurodegenerative diseases.
- Published
- 2015
31. Engineering Poly(ethylene glycol) Particles for Improved Biodistribution
- Author
-
Cui, J, De Rose, R, Alt, K, Alcantara, S, Paterson, BM, Liang, K, Hu, M, Richardson, JJ, Yan, Y, Jeffery, CM, Price, RI, Peter, K, Hagemeyer, CE, Donnelly, PS, Kent, SJ, Caruso, F, Cui, J, De Rose, R, Alt, K, Alcantara, S, Paterson, BM, Liang, K, Hu, M, Richardson, JJ, Yan, Y, Jeffery, CM, Price, RI, Peter, K, Hagemeyer, CE, Donnelly, PS, Kent, SJ, and Caruso, F
- Abstract
We report the engineering of poly(ethylene glycol) (PEG) hydrogel particles using a mesoporous silica (MS) templating method via tuning the PEG molecular weight, particle size, and the presence or absence of the template and investigate the cell association and biodistribution of these particles. An ex vivo assay based on human whole blood that is more sensitive and relevant than traditional cell-line based assays for predicting in vivo circulation behavior is introduced. The association of MS@PEG particles (template present) with granulocytes and monocytes is higher compared with PEG particles (template absent). Increasing the PEG molecular weight (from 10 to 40 kDa) or decreasing the PEG particle size (from 1400 to 150 nm) reduces phagocytic blood cell association of the PEG particles. Mice biodistribution studies show that the PEG particles exhibit extended circulation times (>12 h) compared with the MS@PEG particles and that the retention of smaller PEG particles (150 nm) in blood, when compared with larger PEG particles (>400 nm), is increased at least 4-fold at 12 h after injection. Our findings highlight the influence of unique aspects of polymer hydrogel particles on biological interactions. The reported PEG hydrogel particles represent a new class of polymer carriers with potential biomedical applications.
- Published
- 2015
32. Zn-II(atsm) is protective in amyotrophic lateral sclerosis model mice via a copper delivery mechanism
- Author
-
McAllum, EJ, Roberts, BR, Hickey, JL, Dang, TN, Grubman, A, Donnelly, PS, Liddell, JR, White, AR, Crouch, PJ, McAllum, EJ, Roberts, BR, Hickey, JL, Dang, TN, Grubman, A, Donnelly, PS, Liddell, JR, White, AR, and Crouch, PJ
- Abstract
Mutations in the metalloprotein Cu,Zn-superoxide dismutase (SOD1) cause approximately 20% of familial cases of amyotrophic lateral sclerosis (ALS), a fatal neurodegenerative disease for which effective therapeutics do not yet exist. Transgenic rodent models based on over-expression of mutant SOD1 have been developed and these have provided opportunity to test new therapeutic strategies and to study the mechanisms of mutant SOD1 toxicity. Although the mechanisms of mutant SOD1 toxicity are yet to be fully elucidated, incorrect or incomplete metallation of SOD1 confers abnormal folding, aggregation and biochemical properties, and improving the metallation state of SOD1 provides a viable therapeutic option. The therapeutic effects of delivering copper (Cu) to mutant SOD1 have been demonstrated recently. The aim of the current study was to determine if delivery of zinc (Zn) to SOD1 was also therapeutic. To investigate this, SOD1G37R mice were treated with the metal complex diacetyl-bis(4-methylthiosemicarbazonato)zinc(II) [Zn(II)(atsm)]. Treatment resulted in an improvement in locomotor function and survival of the mice. However, biochemical analysis of spinal cord tissue collected from the mice revealed that the treatment did not increase overall Zn levels in the spinal cord nor the Zn content of SOD1. In contrast, overall levels of Cu in the spinal cord were elevated in the Zn(II)(atsm)-treated SOD1G37R mice and the Cu content of SOD1 was also elevated. Further experiments demonstrated transmetallation of Zn(II)(atsm) in the presence of Cu to form the Cu-analogue Cu(II)(atsm), indicating that the observed therapeutic effects for Zn(II)(atsm) in SOD1G37R mice may in fact be due to in vivo transmetallation and subsequent delivery of Cu.
- Published
- 2015
33. Oral treatment with Cu(II)(atsm) increases mutant SOD1 in vivo but protects motor neurons and improves the phenotype of a transgenic mouse model of amyotrophic lateral sclerosis
- Author
-
Roberts,BR, Lim,NK, McAllum,EJ, Donnelly,PS, Hare,DJ, Doble,PA, Turner,BJ, Price,KA, Lim,SC, Paterson,BM, Hickey,JL, Rhoads,TW, Williams,JR, Kanninen,KM, Hung,LW, Liddell,JR, Grubman,A, Monty,JF, Llanos,RM, Kramer,DR, Mercer,JF, Bush,AI, Masters,CL, Duce,JA, Li,QX, Beckman,JS, Barnham,KJ, White,AR, Crouch,PJ, Roberts,BR, Lim,NK, McAllum,EJ, Donnelly,PS, Hare,DJ, Doble,PA, Turner,BJ, Price,KA, Lim,SC, Paterson,BM, Hickey,JL, Rhoads,TW, Williams,JR, Kanninen,KM, Hung,LW, Liddell,JR, Grubman,A, Monty,JF, Llanos,RM, Kramer,DR, Mercer,JF, Bush,AI, Masters,CL, Duce,JA, Li,QX, Beckman,JS, Barnham,KJ, White,AR, and Crouch,PJ
- Abstract
Mutations in the metallo-protein Cu/Zn-superoxide dismutase (SOD1) cause amyotrophic lateral sclerosis (ALS) in humans and an expression level-dependent phenotype in transgenic rodents. We show that oral treatment with the therapeutic agent diacetyl-bis(4-methylthiosemicarbazonato)copper(II) [Cu(II)(atsm)] increased the concentration of mutant SOD1 (SOD1G37R) in ALS model mice, but paradoxically improved locomotor function and survival of the mice. To determine why the mice with increased levels of mutant SOD1 had an improved phenotype, we analyzed tissues by mass spectrometry. These analyses revealed most SOD1 in the spinal cord tissue of the SOD1G37R mice was Cu deficient. Treating with Cu(II)(atsm) decreased the pool of Cu-deficient SOD1 and increased the pool of fully metallated (holo) SOD1. Tracking isotopically enriched (65)Cu(II)(atsm) confirmed the increase in holo-SOD1 involved transfer of Cu from Cu(II)(atsm) to SOD1, suggesting the improved locomotor function and survival of the Cu(II)(atsm)-treated SOD1G37R mice involved, at least in part, the ability of the compound to improve the Cu content of the mutant SOD1. This was supported by improved survival of SOD1G37R mice that expressed the human gene for the Cu uptake protein CTR1. Improving the metal content of mutant SOD1 in vivo with Cu(II)(atsm) did not decrease levels of misfolded SOD1. These outcomes indicate the metal content of SOD1 may be a greater determinant of the toxicity of the protein in mutant SOD1-associated forms of ALS than the mutations themselves. Improving the metal content of SOD1 therefore represents a valid therapeutic strategy for treating ALS caused by SOD1.
- Published
- 2014
34. X-ray fluorescence imaging reveals subcellular biometal disturbances in a childhood neurodegenerative disorder
- Author
-
Grubman, A, James, SA, James, J, Duncan, C, Volitakis, I, Hickey, JL, Crouch, PJ, Donnelly, PS, Kanninen, KM, Liddell, JR, Cotman, SL, de Jonge, MD, White, AR, Grubman, A, James, SA, James, J, Duncan, C, Volitakis, I, Hickey, JL, Crouch, PJ, Donnelly, PS, Kanninen, KM, Liddell, JR, Cotman, SL, de Jonge, MD, and White, AR
- Abstract
Biometals such as zinc, iron, copper and calcium play key roles in diverse physiological processes in the brain, but can be toxic in excess. A hallmark of neurodegeneration is a failure of homeostatic mechanisms controlling the concentration and distribution of these elements, resulting in overload, deficiency or mislocalization. A major roadblock to understanding the impact of altered biometal homeostasis in neurodegenerative disease is the lack of rapid, specific and sensitive techniques capable of providing quantitative subcellular information on biometal homeostasis in situ. Recent advances in X-ray fluorescence detectors have provided an opportunity to rapidly measure biometal content at subcellular resolution in cell populations using X-ray Fluorescence Microscopy (XFM). We applied this approach to investigate subcellular biometal homeostasis in a cerebellar cell line isolated from a natural mouse model of a childhood neurodegenerative disorder, the CLN6 form of neuronal ceroid lipofuscinosis, commonly known as Batten disease. Despite no global changes to whole cell concentrations of zinc or calcium, XFM revealed significant subcellular mislocalization of these important biological second messengers in cerebellar Cln6nclf (CbCln6nclf ) cells. XFM revealed that nuclear-to-cytoplasmic trafficking of zinc was severely perturbed in diseased cells and the subcellular distribution of calcium was drastically altered in CbCln6nclf cells. Subtle differences in the zinc K-edge X-ray Absorption Near Edge Structure (XANES) spectra of control and CbCln6nclf cells suggested that impaired zinc homeostasis may be associated with an altered ligand set in CbCln6nclf cells. Importantly, a zinc-complex, ZnII(atsm), restored the nuclear-to-cytoplasmic zinc ratios in CbCln6nclf cells via nuclear zinc delivery, and restored the relationship between subcellular zinc and calcium levels to that observed in healthy control cells. ZnII(atsm) treatment also resulted in a reduction in the
- Published
- 2014
35. Deregulation of subcellular biometal homeostasis through loss of the metal transporter, Zip7, in a childhood neurodegenerative disorder
- Author
-
Grubman, A, Lidgerwood, GE, Duncan, C, Bica, L, Tan, J-L, Parker, SJ, Caragounis, A, Meyerowitz, J, Volitakis, I, Moujalled, D, Liddell, JR, Hickey, JL, Horne, M, Longmuir, S, Koistinaho, J, Donnelly, PS, Crouch, PJ, Tammen, I, White, AR, Kanninen, KM, Grubman, A, Lidgerwood, GE, Duncan, C, Bica, L, Tan, J-L, Parker, SJ, Caragounis, A, Meyerowitz, J, Volitakis, I, Moujalled, D, Liddell, JR, Hickey, JL, Horne, M, Longmuir, S, Koistinaho, J, Donnelly, PS, Crouch, PJ, Tammen, I, White, AR, and Kanninen, KM
- Abstract
BACKGROUND: Aberrant biometal metabolism is a key feature of neurodegenerative disorders including Alzheimer's and Parkinson's diseases. Metal modulating compounds are promising therapeutics for neurodegeneration, but their mechanism of action remains poorly understood. Neuronal ceroid lipofuscinoses (NCLs), caused by mutations in CLN genes, are fatal childhood neurodegenerative lysosomal storage diseases without a cure. We previously showed biometal accumulation in ovine and murine models of the CLN6 variant NCL, but the mechanism is unknown. This study extended the concept that alteration of biometal functions is involved in pathology in these disorders, and investigated molecular mechanisms underlying impaired biometal trafficking in CLN6 disease. RESULTS: We observed significant region-specific biometal accumulation and deregulation of metal trafficking pathways prior to disease onset in CLN6 affected sheep. Substantial progressive loss of the ER/Golgi-resident Zn transporter, Zip7, which colocalized with the disease-associated protein, CLN6, may contribute to the subcellular deregulation of biometal homeostasis in NCLs. Importantly, the metal-complex, ZnII(atsm), induced Zip7 upregulation, promoted Zn redistribution and restored Zn-dependent functions in primary mouse Cln6 deficient neurons and astrocytes. CONCLUSIONS: This study demonstrates the central role of the metal transporter, Zip7, in the aberrant biometal metabolism of CLN6 variants of NCL and further highlights the key contribution of deregulated biometal trafficking to the pathology of neurodegenerative diseases. Importantly, our results suggest that ZnII(atsm) may be a candidate for therapeutic trials for NCLs.
- Published
- 2014
36. Rhenium and technetium tricarbonyl complexes of 1,4-substituted pyridyl-1,2,3-triazole bidentate ‘click’ ligands conjugated to a targeting RGD peptide
- Author
-
Connell, TU, Hayne, DJ, Ackermann, U, White, JM, Donnelly, PS, Tochon-Danguy, H, Connell, TU, Hayne, DJ, Ackermann, U, White, JM, Donnelly, PS, and Tochon-Danguy, H
- Abstract
New 1,4‐substituted pyridyl‐1,2,3‐triazole ligands with pendent phenyl isothiocyanate functional groups linked to the heterocycle through a short methylene or longer polyethylene glycol spacers were prepared and conjugated to a peptide containing the arginine–glycine–aspartic acid peptide motif. Rhenium and technetium carbonyl complexes, [M(CO)3Lx(py)]+ (where M = ReI or 99mTcI; Lx = 1,4‐substituted pyridyl‐1,2,3‐triazole ligands and py = pyridine) were prepared. One rhenium complex has been characterized by X‐ray crystallography, and the luminescent properties of [M(CO)3Lx(py)]+ are reported.
- Published
- 2014
37. Neuroprotective Copper Bis(thiosemicarbazonato) Complexes Promote Neurite Elongation
- Author
-
Ginsberg, SD, Bica, L, Liddell, JR, Donnelly, PS, Duncan, C, Caragounis, A, Volitakis, I, Paterson, BM, Cappai, R, Grubman, A, Camakaris, J, Crouch, PJ, White, AR, Ginsberg, SD, Bica, L, Liddell, JR, Donnelly, PS, Duncan, C, Caragounis, A, Volitakis, I, Paterson, BM, Cappai, R, Grubman, A, Camakaris, J, Crouch, PJ, and White, AR
- Abstract
Abnormal biometal homeostasis is a central feature of many neurodegenerative disorders including Alzheimer's disease (AD), Parkinson's disease (PD), and motor neuron disease. Recent studies have shown that metal complexing compounds behaving as ionophores such as clioquinol and PBT2 have robust therapeutic activity in animal models of neurodegenerative disease; however, the mechanism of neuroprotective action remains unclear. These neuroprotective or neurogenerative processes may be related to the delivery or redistribution of biometals, such as copper and zinc, by metal ionophores. To investigate this further, we examined the effect of the bis(thiosemicarbazonato)-copper complex, Cu(II)(gtsm) on neuritogenesis and neurite elongation (neurogenerative outcomes) in PC12 neuronal-related cultures. We found that Cu(II)(gtsm) induced robust neurite elongation in PC12 cells when delivered at concentrations of 25 or 50 nM overnight. Analogous effects were observed with an alternative copper bis(thiosemicarbazonato) complex, Cu(II)(atsm), but at a higher concentration. Induction of neurite elongation by Cu(II)(gtsm) was restricted to neurites within the length range of 75-99 µm with a 2.3-fold increase in numbers of neurites in this length range with 50 nM Cu(II)(gtsm) treatment. The mechanism of neurogenerative action was investigated and revealed that Cu(II)(gtsm) inhibited cellular phosphatase activity. Treatment of cultures with 5 nM FK506 (calcineurin phosphatase inhibitor) resulted in analogous elongation of neurites compared to 50 nM Cu(II)(gtsm), suggesting a potential link between Cu(II)(gtsm)-mediated phosphatase inhibition and neurogenerative outcomes.
- Published
- 2014
38. Oral Treatment with Cu-II(atsm) Increases Mutant SOD1 In Vivo but Protects Motor Neurons and Improves the Phenotype of a Transgenic Mouse Model of Amyotrophic Lateral Sclerosis
- Author
-
Roberts, BR, Lim, NKH, McAllum, EJ, Donnelly, PS, Hare, DJ, Doble, PA, Turner, BJ, Price, KA, Lim, SC, Paterson, BM, Hickey, JL, Rhoads, TW, Williams, JR, Kanninen, KM, Hung, LW, Liddell, JR, Grubman, A, Monty, J-F, Llanos, RM, Kramer, DR, Mercer, JFB, Bush, AI, Masters, CL, Duce, JA, Li, Q-X, Beckman, JS, Barnham, KJ, White, AR, Crouch, PJ, Roberts, BR, Lim, NKH, McAllum, EJ, Donnelly, PS, Hare, DJ, Doble, PA, Turner, BJ, Price, KA, Lim, SC, Paterson, BM, Hickey, JL, Rhoads, TW, Williams, JR, Kanninen, KM, Hung, LW, Liddell, JR, Grubman, A, Monty, J-F, Llanos, RM, Kramer, DR, Mercer, JFB, Bush, AI, Masters, CL, Duce, JA, Li, Q-X, Beckman, JS, Barnham, KJ, White, AR, and Crouch, PJ
- Abstract
Mutations in the metallo-protein Cu/Zn-superoxide dismutase (SOD1) cause amyotrophic lateral sclerosis (ALS) in humans and an expression level-dependent phenotype in transgenic rodents. We show that oral treatment with the therapeutic agent diacetyl-bis(4-methylthiosemicarbazonato)copper(II) [Cu(II)(atsm)] increased the concentration of mutant SOD1 (SOD1G37R) in ALS model mice, but paradoxically improved locomotor function and survival of the mice. To determine why the mice with increased levels of mutant SOD1 had an improved phenotype, we analyzed tissues by mass spectrometry. These analyses revealed most SOD1 in the spinal cord tissue of the SOD1G37R mice was Cu deficient. Treating with Cu(II)(atsm) decreased the pool of Cu-deficient SOD1 and increased the pool of fully metallated (holo) SOD1. Tracking isotopically enriched (65)Cu(II)(atsm) confirmed the increase in holo-SOD1 involved transfer of Cu from Cu(II)(atsm) to SOD1, suggesting the improved locomotor function and survival of the Cu(II)(atsm)-treated SOD1G37R mice involved, at least in part, the ability of the compound to improve the Cu content of the mutant SOD1. This was supported by improved survival of SOD1G37R mice that expressed the human gene for the Cu uptake protein CTR1. Improving the metal content of mutant SOD1 in vivo with Cu(II)(atsm) did not decrease levels of misfolded SOD1. These outcomes indicate the metal content of SOD1 may be a greater determinant of the toxicity of the protein in mutant SOD1-associated forms of ALS than the mutations themselves. Improving the metal content of SOD1 therefore represents a valid therapeutic strategy for treating ALS caused by SOD1.
- Published
- 2014
39. Increasing Intracellular Bioavailable Copper Selectively Targets Prostate Cancer Cells
- Author
-
Cater, MA, Pearson, HB, Wolyniec, K, Klaver, P, Bilandzic, M, Paterson, BM, Bush, AI, Humbert, PO, La Fontaine, S, Donnelly, PS, Haupt, Y, Cater, MA, Pearson, HB, Wolyniec, K, Klaver, P, Bilandzic, M, Paterson, BM, Bush, AI, Humbert, PO, La Fontaine, S, Donnelly, PS, and Haupt, Y
- Abstract
The therapeutic efficacy of two bis(thiosemicarbazonato) copper complexes, glyoxalbis[N4-methylthiosemicarbazonato]Cu(II) [Cu(II)(gtsm)] and diacetylbis[N4-methylthiosemicarbazonato]Cu(II) [Cu(II)(atsm)], for the treatment of prostate cancer was assessed in cell culture and animal models. Distinctively, copper dissociates intracellularly from Cu(II)(gtsm) but is retained by Cu(II)(atsm). We further demonstrated that intracellular H2gtsm [reduced Cu(II)(gtsm)] continues to redistribute copper into a bioavailable (exchangeable) pool. Both Cu(II)(gtsm) and Cu(II)(atsm) selectively kill transformed (hyperplastic and carcinoma) prostate cell lines but, importantly, do not affect the viability of primary prostate epithelial cells. Increasing extracellular copper concentrations enhanced the therapeutic capacity of both Cu(II)(gtsm) and Cu(II)(atsm), and their ligands (H2gtsm and H2atsm) were toxic only toward cancerous prostate cells when combined with copper. Treatment of the Transgenic Adenocarcinoma of Mouse Prostate (TRAMP) model with Cu(II)(gtsm) (2.5 mg/kg) significantly reduced prostate cancer burden (∼70%) and severity (grade), while treatment with Cu(II)(atsm) (30 mg/kg) was ineffective at the given dose. However, Cu(II)(gtsm) caused mild kidney toxicity in the mice, associated primarily with interstitial nephritis and luminal distention. Mechanistically, we demonstrated that Cu(II)(gtsm) inhibits proteasomal chymotrypsin-like activity, a feature further established as being common to copper-ionophores that increase intracellular bioavailable copper. We have demonstrated that increasing intracellular bioavailable copper can selectively kill cancerous prostate cells in vitro and in vivo and have revealed the potential for bis(thiosemicarbazone) copper complexes to be developed as therapeutics for prostate cancer.
- Published
- 2013
40. Increased Zinc and Manganese in Parallel with Neurodegeneration, Synaptic Protein Changes and Activation of Akt/GSK3 Signaling in Ovine CLN6 Neuronal Ceroid Lipofuscinosis
- Author
-
Kahle, PJ, Kanninen, KM, Grubman, A, Meyerowitz, J, Duncan, C, Tan, J-L, Parker, SJ, Crouch, PJ, Paterson, BM, Hickey, JL, Donnelly, PS, Volitakis, I, Tammen, I, Palmer, DN, White, AR, Kahle, PJ, Kanninen, KM, Grubman, A, Meyerowitz, J, Duncan, C, Tan, J-L, Parker, SJ, Crouch, PJ, Paterson, BM, Hickey, JL, Donnelly, PS, Volitakis, I, Tammen, I, Palmer, DN, and White, AR
- Abstract
Mutations in the CLN6 gene cause a variant late infantile form of neuronal ceroid lipofuscinosis (NCL; Batten disease). CLN6 loss leads to disease clinically characterized by vision impairment, motor and cognitive dysfunction, and seizures. Accumulating evidence suggests that alterations in metal homeostasis and cellular signaling pathways are implicated in several neurodegenerative and developmental disorders, yet little is known about their role in the NCLs. To explore the disease mechanisms of CLN6 NCL, metal concentrations and expression of proteins implicated in cellular signaling pathways were assessed in brain tissue from South Hampshire and Merino CLN6 sheep. Analyses revealed increased zinc and manganese concentrations in affected sheep brain in those regions where neuroinflammation and neurodegeneration first occur. Synaptic proteins, the metal-binding protein metallothionein, and the Akt/GSK3 and ERK/MAPK cellular signaling pathways were also altered. These results demonstrate that altered metal concentrations, synaptic protein changes, and aberrant modulation of cellular signaling pathways are characteristic features in the CLN6 ovine form of NCL.
- Published
- 2013
41. An impaired mitochondrial electron transport chain increases retention of the hypoxia imaging agent diacetylbis(4-methylthiosemicarbazonato)copper(II)
- Author
-
Donnelly, PS, Liddell, JR, Lim, S, Paterson, BM, Cater, MA, Savva, MS, Mot, AI, James, JL, Trounce, IA, White, AR, Crouch, PJ, Donnelly, PS, Liddell, JR, Lim, S, Paterson, BM, Cater, MA, Savva, MS, Mot, AI, James, JL, Trounce, IA, White, AR, and Crouch, PJ
- Abstract
Radiolabeled diacetylbis(4-methylthiosemicarbazonato)copper(II) [Cu(II)(atsm)] is an effective positron-emission tomography imaging agent for myocardial ischemia, hypoxic tumors, and brain disorders with regionalized oxidative stress, such as mitochondrial myopathy, encephalopathy, and lactic acidosis with stroke-like episodes (MELAS) and Parkinson's disease. An excessively elevated reductive state is common to these conditions and has been proposed as an important mechanism affecting cellular retention of Cu from Cu(II)(atsm). However, data from whole-cell models to demonstrate this mechanism have not yet been provided. The present study used a unique cell culture model, mitochondrial xenocybrids, to provide whole-cell mechanistic data on cellular retention of Cu from Cu(II)(atsm). Genetic incompatibility between nuclear and mitochondrial encoded subunits of the mitochondrial electron transport chain (ETC) in xenocybrid cells compromises normal function of the ETC. As a consequence of this impairment to the ETC we show xenocybrid cells upregulate glycolytic ATP production and accumulate NADH. Compared to control cells the xenocybrid cells retained more Cu after being treated with Cu(II)(atsm). By transfecting the cells with a metal-responsive element reporter construct the increase in Cu retention was shown to involve a Cu(II)(atsm)-induced increase in intracellular bioavailable Cu specifically within the xenocybrid cells. Parallel experiments using cells grown under hypoxic conditions confirmed that a compromised ETC and elevated NADH levels contribute to increased cellular retention of Cu from Cu(II)(atsm). Using these cell culture models our data demonstrate that compromised ETC function, due to the absence of O(2) as the terminal electron acceptor or dysfunction of individual components of the ETC, is an important determinant in driving the intracellular dissociation of Cu(II)(atsm) that increases cellular retention of the Cu.
- Published
- 2012
42. The challenges of using a copper fluorescent sensor (CS1) to track intracellular distributions of copper in neuronal and glial cells
- Author
-
Price, KA, Hickey, JL, Xiao, Z, Wedd, AG, James, SA, Liddell, JR, Crouch, PJ, White, AR, Donnelly, PS, Price, KA, Hickey, JL, Xiao, Z, Wedd, AG, James, SA, Liddell, JR, Crouch, PJ, White, AR, and Donnelly, PS
- Published
- 2012
43. Inhibition of TDP-43 Accumulation by Bis(thiosemicarbazonato)-Copper Complexes
- Author
-
Kahle, PJ, Parker, SJ, Meyerowitz, J, James, JL, Liddell, JR, Nonaka, T, Hasegawa, M, Kanninen, KM, Lim, S, Paterson, BM, Donnelly, PS, Crouch, PJ, White, AR, Kahle, PJ, Parker, SJ, Meyerowitz, J, James, JL, Liddell, JR, Nonaka, T, Hasegawa, M, Kanninen, KM, Lim, S, Paterson, BM, Donnelly, PS, Crouch, PJ, and White, AR
- Abstract
Amyotrophic lateral sclerosis (ALS) is a progressive, fatal, motor neuron disease with no effective long-term treatment options. Recently, TDP-43 has been identified as a key protein in the pathogenesis of some cases of ALS. Although the role of TDP-43 in motor neuron degeneration is not yet known, TDP-43 has been shown to accumulate in RNA stress granules (SGs) in cell models and in spinal cord tissue from ALS patients. The SG association may be an early pathological change to TDP-43 metabolism and as such a potential target for therapeutic intervention. Accumulation of TDP-43 in SGs induced by inhibition of mitochondrial activity can be inhibited by modulation of cellular kinase activity. We have also found that treatment of cells and animal models of neurodegeneration, including an ALS model, with bioavailable bis(thiosemicarbazonato)copper(II) complexes (Cu(II)(btsc)s) can modulate kinase activity and induce neuroprotective effects. In this study we examined the effect of diacetylbis(-methylthiosemicarbazonato)copper(II) (Cu(II)(atsm)) and glyoxalbis(-methylthiosemicarbazonato)copper(II) (Cu(II)(gtsm)) on TDP-43-positive SGs induced in SH-SY5Y cells in culture. We found that the Cu(II)(btsc)s blocked formation of TDP-43-and human antigen R (HuR)-positive SGs induced by paraquat. The Cu(II)(btsc)s protected neurons from paraquat-mediated cell death. These effects were associated with inhibition of ERK phosphorylation. Co-treatment of cultures with either Cu(II)(atsm) or an ERK inhibitor, PD98059 both prevented ERK activation and blocked formation of TDP-43-and HuR-positive SGs. Cu(II)(atsm) treatment or ERK inhibition also prevented abnormal ubiquitin accumulation in paraquat-treated cells suggesting a link between prolonged ERK activation and abnormal ubiquitin metabolism in paraquat stress and inhibition by Cu. Moreover, Cu(II)(atsm) reduced accumulation of C-terminal (219-414) TDP-43 in transfected SH-SY5Y cells. These results demonstrate that Cu(II)(btsc) com
- Published
- 2012
44. Macrobicyclic Cage Amine Ligands for Copper Radiopharmaceuticals: A Single Bivalent Cage Amine Containing Two Lys(3)-bombesin Targeting Peptides
- Author
-
Ma, MT, Cooper, MS, Paul, RL, Shaw, KP, Karas, JA, Scanlon, D, White, JM, Blower, PJ, Donnelly, PS, Ma, MT, Cooper, MS, Paul, RL, Shaw, KP, Karas, JA, Scanlon, D, White, JM, Blower, PJ, and Donnelly, PS
- Abstract
The synthesis of new cage amine macrobicyclic ligands with pendent carboxylate functional groups designed for application in copper radiopharmaceuticals is described. Reaction of [Cu((NH(2))(2)sar)](2+) (sar = 3,6,10,13,16,19-hexaazabicyclo[6.6.6]icosane) with either succinic or glutaric anhydride results in selective acylation of the primary amine atoms of [Cu((NH(2))(2)sar)](2+) to give derivatives with either one or two aliphatic carboxylate functional groups separated from the cage amine framework by either a four- or five-atom linker. The Cu(II) serves to protect the secondary amine nitrogen atoms from acylation, and can be removed to give the free ligands. The newly appended carboxylate functional groups can be used as sites of attachment for cancer-targeting peptides such as Lys(3)-bombesin. The synthesis of the first dimeric sarcophagine-peptide conjugate, possessing two Lys(3)-bombesin peptides tethered to a single cage amine, is presented. This species has been radiolabeled with copper-64 at ambient temperature and there is minimal dissociation of Cu(II) from the conjugate even after two days of incubation in human serum.
- Published
- 2011
45. Diacetylbis(N(4)-methylthiosemicarbazonato) Copper(II) (Cu-II(atsm)) Protects against Peroxynitrite-induced Nitrosative Damage and Prolongs Survival in Amyotrophic Lateral Sclerosis Mouse Model
- Author
-
Soon, CPW, Donnelly, PS, Turner, BJ, Hung, LW, Crouch, PJ, Sherratt, NA, Tan, J-L, Lim, NK-H, Lam, L, Bica, L, Lim, S, Hickey, JL, Morizzi, J, Powell, A, Finkelstein, DI, Culvenor, JG, Masters, CL, Duce, J, White, AR, Barnham, KJ, Li, Q-X, Soon, CPW, Donnelly, PS, Turner, BJ, Hung, LW, Crouch, PJ, Sherratt, NA, Tan, J-L, Lim, NK-H, Lam, L, Bica, L, Lim, S, Hickey, JL, Morizzi, J, Powell, A, Finkelstein, DI, Culvenor, JG, Masters, CL, Duce, J, White, AR, Barnham, KJ, and Li, Q-X
- Abstract
Amyotrophic lateral sclerosis (ALS) is a progressive paralyzing disease characterized by tissue oxidative damage and motor neuron degeneration. This study investigated the in vivo effect of diacetylbis(N(4)-methylthiosemicarbazonato) copper(II) (CuII(atsm)), which is an orally bioavailable, blood-brain barrier-permeable complex. In vitro the compound inhibits the action of peroxynitrite on Cu,Zn-superoxide dismutase (SOD1) and subsequent nitration of cellular proteins. Oral treatment of transgenic SOD1G93A mice with CuII(atsm) at presymptomatic and symptomatic ages was performed. The mice were examined for improvement in lifespan and motor function, as well as histological and biochemical changes to key disease markers. Systemic treatment of SOD1G93A mice significantly delayed onset of paralysis and prolonged lifespan, even when administered to symptomatic animals. Consistent with the properties of this compound, treated mice had reduced protein nitration and carbonylation, as well as increased antioxidant activity in spinal cord. Treatment also significantly preserved motor neurons and attenuated astrocyte and microglial activation in mice. Furthermore, CuII(atsm) prevented the accumulation of abnormally phosphorylated and fragmented TAR DNA-binding protein-43 (TDP-43) in spinal cord, a protein pivotal to the development of ALS. CuII(atsm) therefore represents a potential new class of neuroprotective agents targeting multiple major disease pathways of motor neurons with therapeutic potential for ALS.
- Published
- 2011
46. Cellular Up-regulation of Nedd4 Family Interacting Protein 1 (Ndfip1) using Low Levels of Bioactive Cobalt Complexes
- Author
-
Schieber, C, Howitt, J, Putz, U, White, JM, Parish, CL, Donnelly, PS, Tan, S-S, Schieber, C, Howitt, J, Putz, U, White, JM, Parish, CL, Donnelly, PS, and Tan, S-S
- Abstract
The delivery of metal ions using cell membrane-permeable metal complexes represents a method for activating cellular pathways. Here, we report the synthesis and characterization of new [Co(III)(salen)(acac)] complexes capable of up-regulating the ubiquitin ligase adaptor protein Ndfip1. Ndfip1 is a neuroprotective protein that is up-regulated in the brain after injury and functions in combination with Nedd4 ligases to ubiquitinate harmful proteins for removal. We previously showed that Ndfip1 can be increased in human neurons using CoCl(2) that is toxic at high concentration. Here we demonstrate a similar effect can be achieved by low concentrations of synthetic Co(III) complexes that are non-toxic and designed to be activated following cellular entry. Activation is achieved by intracellular reduction of Co(III) to Co(II) leading to release of Co(II) ions for Ndfip1 up-regulation. The cellular benefit of Ndfip1 up-regulation by Co(III) complexes includes demonstrable protection against cell death in SH-SY5Y cells during stress. In vivo, focal delivery of Co(III) complexes into the adult mouse brain was observed to up-regulate Ndfip1 in neurons. These results demonstrate that a cellular response pathway can be advantageously manipulated by chemical modification of metal complexes, and represents a significant step of harnessing low concentration metal complexes for therapeutic benefit.
- Published
- 2011
47. Versatile New Bis(thiosemicarbazone) Bifunctional Chelators: Synthesis, Conjugation to Bombesin(7-14)-NH2, and Copper-64 Radiolabeling
- Author
-
Paterson, BM, Karas, JA, Scanlon, DB, White, JM, Donnelly, PS, Paterson, BM, Karas, JA, Scanlon, DB, White, JM, and Donnelly, PS
- Abstract
New bifunctional derivatives of diacetyl-bis(4-methylthiosemicarbazone) (H(2)atsm) have been prepared by a selective transamination reaction of a new dissymmetric bis(thiosemicarbazone) precursor H(2)L(1). The new derivatives contain an aliphatic carboxylic acid (H(2)L(2) and H(2)L(3)), t-butyl carbamate (H(2)L(4)), or ammonium ion (H(2)L(5)) functional group. The new ligands and copper(II) complexes have been characterized by NMR spectroscopy, mass spectrometry, and microanalysis. The complex Cu(II)(L(4)) was structurally characterized by X-ray crystallography and shows the metal center to be in an N(2)S(2) distorted square planar coordination geometry. Electrochemical measurements show that the copper(II) complexes undergo a reversible reduction attributable to a Cu(II)/Cu(I) process. The ligands and the copper(II) complexes featuring a carboxylic acid functional group have been conjugated to the tumor targeting peptide bombesin(7-14)-NH(2). The bifunctional peptide conjugates were radiolabeled with copper-64 in the interest of developing new positron emission tomography (PET) imaging agents. The conjugates were radiolabeled with copper-64 rapidly in high radiochemical purity (>95%) at room temperature under mild conditions and were stable in a cysteine and histidine challenge study.
- Published
- 2010
48. Membrane-targeted strategies for modulating APP and A beta-mediated toxicity
- Author
-
Price, KA, Crouch, PJ, Donnelly, PS, Masters, CL, White, AR, Curtain, CC, Price, KA, Crouch, PJ, Donnelly, PS, Masters, CL, White, AR, and Curtain, CC
- Abstract
Alzheimer's disease (AD) is a progressive neurodegenerative disease characterized by numerous pathological features including the accumulation of neurotoxic amyloid-beta (Abeta) peptide. There is currently no effective therapy for AD, but the development of therapeutic strategies that target the cell membrane is gaining increased interest. The amyloid precursor protein (APP) from which Abeta is formed is a membrane-bound protein, and Abeta production and toxicity are both membrane mediated events. This review describes the critical role of cell membranes in AD with particular emphasis on how the composition and structure of the membrane and its specialized regions may influence toxic or benign Abeta/APP pathways in AD. The putative role of copper (Cu) in AD is also discussed, and we highlight how targeting the cell membrane with Cu complexes has therapeutic potential in AD.
- Published
- 2009
49. Bis(thiosemicarbazonato) Cu-64 complexes for positron emission tomography imaging of Alzheimer's disease.
- Author
-
Fodero-Tavoletti MT, Villemagne VL, Paterson BM, White AR, Li QX, Camakaris J, O'Keefe G, Cappai R, Barnham KJ, Donnelly PS, Fodero-Tavoletti, Michelle T, Villemagne, Victor L, Paterson, Brett M, White, Anthony R, Li, Qiao-Xin, Camakaris, James, O'Keefe, Graeme, Cappai, Roberto, Barnham, Kevin J, and Donnelly, Paul S
- Abstract
A bis (thiosemicarbazonato) complex radiolabeled with positron emitting Cu-64 can be used for a new and alternative method for the non-invasive diagnosis of Alzheimer's disease using positron emission tomography (PET). Most imaging agents being investigated for the diagnosis of Alzheimer's disease target amyloid-beta plaque burden but our new approach highlights altered copper homeostasis. This approach has the potential to offer complementary information to other diagnostic procedures that elucidate plaque burden. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF
50. Increased zinc and manganese in parallel with neurodegeneration, synaptic protein changes and activation of Akt/GSK3 signaling in ovine CLN6 neuronal ceroid lipofuscinosis
- Author
-
Kanninen, KM, Grubman, A, Meyerowitz, J, Duncan, C, Tan, J-L, Parker, SJ, Crouch, PJ, Paterson, BM, Hickey, JL, Donnelly, PS, Volitakis, I, Tammen, I, Palmer, DN, and White, AR
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.