Monoclonal antibody specific for the transverse tubular membrane of skeletal muscle activates the dihydropyridine-sensitive Ca2+ channel.

Malouf, Nadia N.; Coronado, Roberto; Mcmahon, Debra K.; Meissner, Gerhard; Gillespie, G. Yancey

doi:10.1073/pnas.84.14.5019

Cited by 12 publications

(4 citation statements)

References 29 publications

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“…mHK2-ASO1 and mHK2-ASO2 suppressed mouse HK2 expression in mouse multiple myeloma P3 Â 63Ag (P3) cells (20) without affecting HK1 expression ( Supplementary Fig. S7C).…”

Section: Hk2-aso/dpi/per and Hk2-aso/met/per Combinations Are Toleratmentioning

confidence: 95%

An HK2 Antisense Oligonucleotide Induces Synthetic Lethality in HK1−HK2+ Multiple Myeloma

Zhou

Doh

et al. 2019

Cancer Research

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A synthetically lethal precision medicine therapy for HK1-HK2+ multiple myeloma using an HK2 antisense oligonucleotide, metformin, and perhexiline. Most normal tissues Tolerated Tolerated HK1 + HK2 + multiple myeloma HK 2-A SO Me tfo rmi n Per hex ilin e HK1-HK2 + multiple myeloma Glucose-6-P Glucose-6-P Glucose Glucose HK1 HK2 HK2 Synthetically lethal Although the majority of adult tissues express only hexokinase 1 (HK1) for glycolysis, most cancers express hexokinase 2 (HK2) and many coexpress HK1 and HK2. In contrast to HK1 þ HK2 þ cancers, HK1 À HK2 þ cancer subsets are sensitive to cytostasis induced by HK2 shRNA knockdown and are also sensitive to synthetic lethality in response to the combination of HK2 shRNA knockdown, an oxidative phosphorylation (OXPHOS) inhibitor diphenyleneiodonium (DPI), and a fatty acid oxidation (FAO) inhibitor perhexiline (PER). The majority of human multiple myeloma cell lines are HK1 À HK2 þ. Here we describe an antisense oligonucleotide (ASO) directed against human HK2 (HK2-ASO1), which suppressed HK2 expression in human multiple myeloma cell cultures and human multiple myeloma mouse xenograft models. The HK2-ASO1/DPI/PER triple-combination achieved synthetic lethality in multiple myeloma cells in culture and prevented HK1 À HK2 þ multiple myeloma tumor xenograft progression. DPI was replaceable by the FDA-approved OXPHOS inhibitor metformin (MET), both for synthetic lethality in culture and for inhibition of tumor xenograft progression. In addition, we used an ASO targeting murine HK2 (mHK2-ASO1) to validate the safety of mHK2-ASO1/MET/PER combination therapy in mice bearing murine multiple myeloma tumors. HK2-ASO1 is the first agent that shows selective HK2 inhibition and therapeutic efficacy in cell culture and in animal models, supporting clinical development of this synthetically lethal combination as a therapy for HK1 À HK2 þ multiple myeloma. Significance: A first-in-class HK2 antisense oligonucleotide suppresses HK2 expression in cell culture and in in vivo, presenting an effective, tolerated combination therapy for preventing progression of HK1 À HK2 þ multiple myeloma tumors.

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“…mHK2-ASO1 and mHK2-ASO2 suppressed mouse HK2 expression in mouse multiple myeloma P3 Â 63Ag (P3) cells (20) without affecting HK1 expression ( Supplementary Fig. S7C).…”

Section: Hk2-aso/dpi/per and Hk2-aso/met/per Combinations Are Toleratmentioning

confidence: 95%

An HK2 Antisense Oligonucleotide Induces Synthetic Lethality in HK1−HK2+ Multiple Myeloma

Zhou

Doh

et al. 2019

Cancer Research

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“…Further evidence that the phosphorylatable 165 kDa (o~) peptide is associated with the DHP receptor came from studies showing that two different monoclonal antibodies (Leung, Imagawa & Campbell, 1987;Morton & Froehner, 1987) and a polyclonal antibody (Takahashi & Catterall, 1987) were able to specifically immunoprecipitate the [3H]PN200-100 labeled receptors from solubilized preparations and recognized the 165 kDa peptide in Western blots. A third monoclonal antibody, which activates DHP-sensitive Ca channels in skeletal muscle membranes, may also recognize the I65 kDa peptide (Malouf et al, 1987).…”

Section: Purification Of the Dihydropyridine And Phenylalkylamine-senmentioning

confidence: 99%

Calcium channels: Molecular pharmacology, structure and regulation

1988

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“…Specific antibodies have been used to explore several functional and structural properties of ionic channels (Meiri et al, 1986;Malouf et al, 1987;Morton et al, 1988;Fitzpatrick et al, 1988).…”

Section: Introductionmentioning

confidence: 99%

Antibodies as probes for ligand gating of single sarcoplasmic reticulum Ca2+-release channels

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Mejía-Alvarez

Zorzato

et al. 1991

Biochemical Journal

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A large (565 kDa) junctional sarcoplasmic reticulum (SR) protein, the ryanodine receptor (RYR), may play both a structural and a functional role in the mechanism of skeletal muscle excitation-contraction coupling. Recently, the primary amino acid sequence of the RYR has been elucidated. In this paper, we introduce an immunological approach to examine the functional (electrophysiological) properties of the RYR when it is incorporated into planar lipid bilayers. The effects of two polyclonal antibodies against the SR junctional face membrane (JFM) and the RYR (anti-JFM and anti-RYR) were tested on the single-channel gating properties of the RYR SR Ca2(+)-release channel. Junctional SR vesicles were fused into planar lipid bilayers in solutions containing caesium salts. Solutions were designed to minimize the background conductances of the SR K+ and Cl- channels. Three actions of the anti-JFM antibody were distinguished on the basis of single-channel gating and conductance. The anti-RYR antibody had a single action, a simultaneous decrease in single-channel open probability (Po) and conductance. Both antibodies appear to alter single-channel gating by disrupting the Ca2(+)-activation mechanism of the channel. Anti-RYR-antibody-induced gating abnormalities were reversed by ATP, although the ATP-re-activated channels had altered gating kinetics. Two antigenic regions, recognizing the anti-RYR antibody, in the C-terminal end of the RYR primary amino acid sequence contain or are closely associated with putative ligand (Ca2+ and ATP)-binding sites identified previously. Our results demonstrate (1) that the antibodies induced abnormal gating (decreased open probability and stabilization of subconducting states) of SR release channels, and (2) that abnormal gating is not associated with physical obstruction or alteration of the conduction pathway. Thus antibodies directed at specific regions of the RYR (e.g. putative ligand-binding sites) can be used as effective probes with which to study the structural and functional properties of the SR Ca2(+)-release channel gating at the single-channel level.

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Monoclonal antibody specific for the transverse tubular membrane of skeletal muscle activates the dihydropyridine-sensitive Ca2+ channel.

Abstract: In skeletal muscle, dihydropyridine receptors and dihydropyridine-sensitive

Cited by 12 publications

References 29 publications

An HK2 Antisense Oligonucleotide Induces Synthetic Lethality in HK1−HK2+ Multiple Myeloma

An HK2 Antisense Oligonucleotide Induces Synthetic Lethality in HK1−HK2+ Multiple Myeloma

Calcium channels: Molecular pharmacology, structure and regulation

Antibodies as probes for ligand gating of single sarcoplasmic reticulum Ca2+-release channels

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