University of Iowa Research Foundation

Other

Miles Inc.

University of Minnesota

Washington University

The Rockefeller University

Sibia Neurosciences, Inc.

The Salk Institute Biotechnology Industrial Associates, Inc.

Univ. of Iowa Research Foundation

Kevin P Campbell

Steven D Kahl

James M Ervasti

Roger Williamson

Franck Duclos

Volker Straub

Yoshihide Sunada

Michael Miller Harpold

Steven B Ellis

Daniel Yonil Jung

Michael D Henry

James R Mickelson

Steven L Roberds

Kay Ohlendieck

Cynthia J Leveille

C Michael Knudson

Charles F Louis

Yvonne M Kobayashi

Leland E Lim

Mitchell G Gaver

Michel Fardeau

Robert W Crawford

John McPherson

Fernando M Tome

Scott David Jay

Vincent A Fischetti

Beverly L Davidson

Jeffrey Allen Campbell

Terence Lewis

Kyle B Campbell

Thomas Judson Campbell

Michael B Cohen

Wayne N Frankel

Richard D Anderson

Michael Oldstone

Madeleine Durbeej

Ronald Cohn

Valerie Allamand

Mustafa Salih

Hiroki Yamada

Verity A Letts

Wei Cao

Toshiaki Imagawa

Rita Barresi

Gloria Biddlecome

Albert T Leung

Ricardo Felix

Oxana Ibraghimov-Beskrovnaya

Anura Rambukkana

Kiichiro Matsumura

Philip Bentley

Kathleen H Holt

John D Windass

Peter McPherson

Derrick R Witcher

Oxana B Ibraghimov

Charles E Jackson

Rachelle Crosbie

Jacques S Beckmann

Odile Broux

Connie Lebakken

Federica Piccolo

Marc Jeanpierre

Ramon Coral

Jean-Claude Kaplan

Disclosed are pharmaceutical compositions and methods for treating or preventing muscle diseases or the symptoms thereof. The compositions typically include and the methods typically utilize phosphodiesterase type 5A inhibitors.

Inhibitors of phosphodiesterase type 5A for reducing skeletal muscle fatigue, edema, and damage in a patient having muscle fatigue due to increased age or exercise

Inhibitors of phosphodiesterase type 5A for treating or preventing muscle disease or the symptoms thereof in a patient

Disclosed is a method for the prevention and/or treatment of muscle degeneration. In this method, a subject recognized as having muscle degeneration is treated with a composition effective to increase functional glycosylation of &#x3b1;-dystroglycan in an affected tissue in the subject. Functional glycosylation is to be increased to an extent wherein the binding of &#x3b1;-dystroglycan to its ligands in the affected tissue is rescued to levels substantially similar to those in an evenly matched tissue unaffected by degeneration. One effective means for increasing functional glycosylation of &#x3b1;-dystroglycan in a subject includes increasing glycosyltransferase activity, such as LARGE or LARGE2 activity, in the muscle of the subject. Therapeutic glycosylated peptide compositions are also provided.

Increasing functional glycosylation of alpha-dystroglycan in the treatment of muscle degeneration

A conveyor system is provided to limit the vertical fall experienced by a product transported along the conveyor and falling into a collection bin. A sensor in a fixed position relative to the product conveying plane of the conveyor measures product collection height in a collection bin to produce an output signal. A controller dynamically varies the angle between the conveyance plane and the collection bin base plane and the vertical displacement therebetween within a preselected range in order to minimize product damage associated with different fall trajectories from the conveyor into the bin for product. Either the conveyor or the collection bin is pivoted under the command of the controller to accomplish the desired function.

Conveyor system for minimizing product damage during collection

Disclosed is a method for diagnosing the tumorigenic grade of a malignant tissue. The method entails determining the amount of dystroglycan protein of the malignant tissue relative to a standard. Suitable methods for determining the amount of dystroglycan protein of the tissue are provided, and include measuring the amount of mRNA transcripts which encode dystroglycan, and also performing western blot analysis or immunofluorescence analysis on the tissue components to detect &agr;-dystroglycan or &bgr;-dystroglycan. An antibody probe which binds specifically to the C-terminus of &bgr;-dystroglycan, is provided. This method is applicable to human malignant tissue, especially adenocarcinoma, and preferably prostate or mammary adenocarcinoma. This method can also be applied to the detection of a cancerous disease state in a tissue of a patient, with a decreased level of dystroglycan protein being indicative of the presence of cancer. Also disclosed is a method for determining the prognosis of a patient afflicted with a malignancy by determining the expression level of the dystroglycan gene in a tissue sample of the malignancy, and comparing the expression level to a standard, with a decreased level of dystroglycan expression being indicative of unfavorable prognosis. A method for identifying an individual at risk for the development of cancer, or an individual at risk for the recurrence of cancer after treatment, is also disclosed. Similarly, a method for identifying individuals at risk for developing cancer by screening for mutations in the dystroglycan genes of the individual is also provided. One such mutation is the allelic loss of human chromosome 3p21.

Evaluation of adenocarcinoma of the prostate and breast using anti-dystroglycan antibodies

Disclosed are mammalian nucleic acid sequences encoding a neuronal-specific subunit of a voltage-gated calcium channel. Specifically disclosed are &ggr; , &ggr; and &ggr; subunits. In other aspects, the disclosure relates to expression vectors which encode neuronal-specific subunits, as well as cells containing such vectors. In other aspects, the disclosure relates to antigenic fusion proteins comprising at least a portion of a mammalian neuronal-specific subunit of a voltage-gated calcium channel. Such fusion proteins are useful, for example, in the production of antibodies specifically reactive with the subunits of the invention. The nucleic acid sequences of the invention find application, for example, in screening for compounds which modulate the activity of neuronal voltage-gated calcium channels and also in diagnostic methods for diagnosing the autoimmune disease Lambert-Eaton Syndrome, as well as diagnosing defects in &ggr; subunit genes of a patient with a neuronal disease such as epilepsy. An additional application of the nucleic acid sequences of the invention is in therapeutic methods of treatment for such disorders.

Genes encoding neuronal voltage-gated calcium channel gamma subunits

The present invention relates to the discovery that the &agr;-dystroglycan receptor is required for,entry into cells, assays for high throughput screening of drugs for use in treatments against leprosy, and methods for studying the role of the receptor in neurodegenerative and musculodegenerative diseases, and the like.

Receptor for Mycobacterium leprae and methods of use thereof

Disclosed is a method for treating a patient suffering from the disease sarcoglycan-deficient limb-girdle muscular dystrophy by gene replacement therapy. Sarcoglycan gene replacement therapy produces extensive long-term expression of the sarcoglycan species which restores the entire sarcoglycan complex, results in the stable association of alph&agr;-dystroglycan with the sarcolemma, and eliminates the morphological markers of limb-girdle muscular dystrophy. In another aspect, the invention relates to a method for determining a specific defective sarcoglycan species in the tissue of a patient. The method involves culture of muscle cells obtained from the patient, and the independent introduction of expression vectors encoding each of the sarcoglycan species, &agr;, &bgr;, &ggr;, and &dgr;, into the cultured cells with subsequent assaying for restoration of the dystrophin-glycoprotein complex. In another aspect, the invention relates to a mouse, and cells derived therefrom, homozygous for a disrupted &agr;-sarcoglycan gene. The disruption prevents the synthesis of functional &agr;-sarcoglycan in cells of the mouse and results in the mutant mouse having no detectable sarcospan, &bgr;-, &ggr;-, &dgr;-sarcoglycan, and reduced &agr;-dystroglycan in the sarcolemma of skeletal and cardiac muscles, and a reduction of dystrophin in skeletal muscle, when compared to tissue of a mouse lacking a disrupted &agr;-sarcoglycan gene. In another aspect, the invention relates to methods for screening for therapeutic agents useful in the treatment of sarcoglycan-deficient limb-girdle muscular dystrophy. The methods involve administering a candidate therapeutic agent to a mouse, or cells derived therefrom, and assaying for therapeutic effects on the mouse or cells, with the determination of therapeutic effects being a reduction or reversal in disease progression, or a restoration of the dystroglycan complex.

Gene replacement therapy for muscular dystrophy

Disclosed herein is a substantially pure nucleic acid sequence encoding a mammalian 35 kDa non-dystrophin component (&dgr;-sarcoglycan) of the dystrophin-glycoprotein complex. Also disclosed are the amino acid sequence and an immunogenic peptide of &dgr;-sarcoglycan. The peptide when used to immunize a mammal, stimulates the production of antibodies which bind specifically to the &dgr;-sarcoglycan. Methods to identify mutations in the &dgr;-sarcoglycan gene associated with autosomal recessive limb-girdle muscular dystrophy are also disclosed. The identification of such mutations enables the design of nucleic acid probes which hybridize specifically to a mutant form of &dgr;-sarcoglycan, or the complement thereof, but not to the DNA of the wild-type form of the gene (or the complement thereof), under stringent hybridization conditions. Such probes are useful, for example, in connection with the diagnosis of autosomal recessive limb-girdle muscular dystrophy. In addition, the identification of such mutations enables the diagnosis of autosomal recessive limb-girdle muscular dystrophy through the use of direct DNA sequencing techniques.

&dgr;-sarcoglycan antibodies

Disclosed is a transgenic knockout mouse whose genome has a homozygous disruption in its endogenous sarcospan gene, wherein the disruption prevents the synthesis of functional sarcospan in cells of the mouse. The mouse is characterized as exhibiting from 1.4 to 6.8 fold larger epididymal fat pad deposits as compared to the epididymal fat pad deposits of a wild type mouse. Methods for production of the mouse are presented. Also disclosed are cells derived from the transgenic knockout mouse. The mouse can be used in a method for identifying therapeutic agents for the treatment of an individual diagnosed with a metabolic disorder associated with a reduction or loss of expression of wild-type sarcospan. An example of such a disorder is weight gain in the individual associated with a reduction or loss of expression of wild-type sarcospan. These specific methods are also provided.

Sarcospan-deficient mouse as a model for clinical disorders associated with sarcospan mutations

Disclosed is a mouse, cells derived therefrom, and methods for using the mouse, the mouse being homozygous for a disrupted &dgr;-sarcoglycan gene, the disruption in the gene having been introduced into the mouse or an ancestor of the mouse at an embryonic stage. The disruption prevents the synthesis of functional &dgr;-sarcoglycan in cells of the mouse and results in the mouse having a reduced amount of &bgr;- and &egr;-sarcoglycan and sarcospan, and a disruption of the sarcoglycan-sarcospan complex in smooth muscle of the mouse. Also disclosed is a mouse, cells derived therefrom, and methods for using the mouse, the mouse being homozygous for a disrupted &bgr;-sarcoglycan gene, the disruption in the gene having been introduced into the mouse or an ancestor of the mouse at an embryonic stage. The disruption prevents the synthesis of functional &bgr;-sarcoglycan in cells of the mouse and results in the mouse having a reduced amount of &dgr;-and &egr;-sarcoglycan and sarcospan and &agr;-dystroglycan in smooth muscle of the mouse.

Pathogenesis of cardiomyopathy

Disclosed are compositions and methods for aiding in the diagnosis of congenital muscular dystrophy associated with in-frame deletion in the laminin-2 .alpha.2 polypeptide chain in an individual. In a preferred diagnostic method embodiment, an experimental muscle tissue sample is provided from the individual and treated if necessary to render components available for antibody binding. The components of the sample are then separated on the basis of molecular weight. The separated protein components are then transferred to a solid support while maintaining the relative positions established in separation step. The transferred components are then stained with an affinity reagent which is known to bind to a C-terminal domain of the laminin-2 .alpha.2 polypeptide chain. Individual afflicted with congenital muscular dystrophy associated with in-frame deletion in the laminin-2 .alpha.2 polypeptide chain on the basis of positive staining in combination with reduced molecular weight of the laminin-2 .alpha.2 polypeptide chain relative to the wild-type laminin-2 .alpha.2 polypeptide chain. A preferred composition is a nucleic acid probe for the detection of merosin deletion-type congenital muscular dystrophy. The preferred nucleic acid probe is characterized by the ability to bind specifically to a mutant merosin nucleic acid sequence, the mutant merosin nucleic acid sequence comprising a T to C substitution at position 3973 +2 of the consensus donor splice site of exon 25.

Method for aiding in the diagnosis of in-frame deletion type congenital

Disclosed is a method for inhibiting the binding of an arenavirus to a cellular receptor. The method involves providing, in soluble form, a reagent comprising .alpha.-dystroglycan or a portion thereof, the reagent being characterized by the ability to bind to the arenavirus thereby inhibiting the binding of the arenavirus to the cellular receptor. The reagent is contacted with an arenavirus particle prior to infection of a cell by the arenavirus particle. Also disclosed are methods for treating an arenavirus infection in a patient and preventing an arenavirus infection in an individual at risk. These methods involve providing a therapeutic composition comprising .alpha.-dystroglycan or a portion thereof which is characterized by the ability to bind to arenaviruses, thereby inhibiting the binding of arenaviruses to a cellular receptor; and administering the composition to the patient or individual at risk. Arenaviruses to which the methods of the present invention apply include, without limitation, Lymphocyte Choriomeningitis Virus, Lassa fever virus, Mobala, and Oliveros. In another aspect, the disclosure relates to an embryonic stem cell line, and cells derived therefrom, which is homozygous for a disrupted dystroglycan gene, wherein the disruption prevents the synthesis of functional dystroglycan in the cells. Applications of the dystroglycan null embryonic stem cells include producing dystroglycan or a portion thereof in the cells and also for identifying portions of dystroglycan necessary for arenavirus infection. Also disclosed is a method for identifying antiviral compounds which interfere specifically with the binding of arenavirus and .alpha.-dystroglycan, comprising providing a binding assay system for the determination of binding of arenavirus and .alpha.-dystroglycan. The candidate antiviral compounds are introduced into the binding assay system and antiviral compounds which substantially inhibit binding of arenavirus to .alpha.-dystroglycan are identified.

Arenavirus receptor and methods of use

Disclosed is a method for aiding in the diagnosis of merosin deficiency-type congenital muscular dystrophy (CMD). The method is based on the discovery of a previously unidentified form of CMD which is characterized by a substantial reduction in the levels of merosin in skeletal muscle tissue containing normal levels of dystrophin and dystrophin-associated proteins.

Merosin deficiency-type congenital muscular dystrophy

Disclosed herein is a substantially pure nucleic acid sequence encoding a mammalian 35 kDa non-dystrophin component (.delta.-sarcoglycan) of the dystrophin-glycoprotein complex. Also disclosed are the amino acid sequence and an immunogenic peptide of .delta.-sarcoglycan. The peptide when used to immunize a mammal, stimulates the production of antibodies which bind specifically to the .delta.-sarcoglycan. Methods to identify mutations in the .delta.-sarcoglycan gene associated with autosomal recessive limb-girdle muscular dystrophy are also disclosed. The identification of such mutations enables the design of nucleic acid probes which hybridize specifically to a mutant form of .delta.-sarcoglycan, or the complement thereof, but not to the DNA of the wild-type form of the gene (or the complement thereof), under stringent hybridization conditions. Such probes are useful, for example, in connection with the diagnosis of autosomal recessive limb-girdle muscular dystrophy. In addition, the identification of such mutations enables the diagnosis of autosomal recessive limb-girdle muscular dystrophy through the use of direct DNA sequencing techniques.

.delta.-sarcoglycan nucleic acid sequences

Growing community of inventors

Iowa City, IA, United States of America

Kevin P Campbell