Materials and methods for treatment of amyotrophic lateral sclerosis and/or frontal temporal lobular degeneration
AssigneeCrispr Therapeutics AG
InventorsChad Albert Cowan, Ami Meda Kabadi, Ante Sven Lundberg
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Abstract
The present application provides materials and methods for treating a patient with Amyotrophic Lateral Sclerosis (ALS) and/or Frontaltemporal Lobular Degeneration (FTLD), both ex vivo and in vivo. In addition, the present application provides materials and methods for editing to modulate the expression, function or activity of the C9ORF72 gene in a cell by genome editing.
Claims80
A method for editing the C9ORF72 gene in a human cell by genome editing comprising introducing into the cell one or more deoxyribonucleic acid (DNA) endonucleases to effect one or more single-strand breaks (SSBs) or double-strand breaks (DSBs) within or near the C9ORF72 gene or other DNA sequences that encode regulatory elements of the C9ORF72 gene that results in permanent deletion, insertion, or correction of the expanded hexanucleotide repeat within or near the C9ORF72 gene and results in restoration of C9orf72 protein activity.
A method for inserting the C9ORF72 gene in a human cell by genome editing, the method comprising the step of:
- introducing into the human cell one or more deoxyribonucleic acid (DNA) endonucleases to effect one or more single-strand breaks (SSBs) or double-strand breaks (DSBs) within or near a safe harbor locus that results in a permanent insertion of the C9ORF72 gene or minigene, and results in restoration of the C9orf72 protein activity.
An ex vivo method for treating a patient with amyotrophic lateral sclerosis (ALS) or frontotemporal lobular dementia (FTLD) comprising the steps of:
- i) creating a patient specific induced pluripotent stem cell (iPSC);
- ii) editing within or near the C9ORF72 gene of the iPSC or other DNA sequences that encode regulatory elements of the C9ORF72 gene of the iPSC, or editing within or near a safe harbor locus of the iPSC;
- iii) differentiating the genome edited iPSC into a hematopoietic progenitor cell, a neural progenitor cell, or neural cell;
- and
- iv) implanting the hematopoietic progenitor cell, neural progenitor cell, or neural cell into the patient.
An ex vivo method for treating a patient with amyotrophic lateral sclerosis (ALS) or frontotemporal lobular dementia (FTLD) comprising the steps of:
- i) isolating a white blood cell from the patient;
- ii) editing within or near the C9ORF72 gene or other DNA sequences that encode regulatory elements of the C9ORF72 gene of the white blood cell, or editing within or near a safe harbor locus of the white blood cell;
- and
- iii) implanting the edited white blood cell into the patient.
An ex vivo method for treating a patient with amyotrophic lateral sclerosis (ALS) or frontotemporal lobular dementia (FTLD) comprising the steps of:
- i) optionally, performing a biopsy of the patient's bone marrow;
- ii) isolating a mesenchymal stem cell;
- iii) editing within or near the C9ORF72 gene of the stem cell or other DNA sequences that encode regulatory elements of the C9ORF72 gene of the stem cell, or editing within or near a safe harbor locus of the stem cell;
- iv) differentiating the stem cell into a hematopoietic progenitor cell, neural progenitor cell, or neural cell;
- and
- v) implanting the hematopoietic progenitor cell, neural progenitor cell, or neural cell into the patient.
An ex vivo method for treating a patient with amyotrophic lateral sclerosis (ALS) or frontotemporal lobular dementia (FTLD) comprising the steps of:
- i) optionally, treating the patient with granulocyte colony stimulating factor (GCSF);
- ii) isolating a hematopoietic progenitor cell from the patient;
- iii) editing within or near the C9ORF72 gene of the hematopoietic progenitor cell or other DNA sequences that encode regulatory elements of the C9ORF72 gene of the hematopoietic progenitor cell, or editing within or near a safe harbor locus of the hematopoietic progenitor cell;
- and
- iv) implanting the cell into the patient.
An in vivo method for treating a patient with amyotrophic lateral sclerosis (ALS) or frontotemporal lobular dementia (FTLD) comprising the step of editing within or near the C9ORF72 gene in a cell of the patient or other DNA sequences that encode regulatory elements of the C9ORF72 gene, or editing within or near a safe harbor locus.
The method of any one of the preceding claims, wherein the method further comprises introducing into the cell one or more guide ribonucleic acids (gRNAs).
The method of any one of the preceding claims, wherein the method further comprises introducing into the cell a polynucleotide donor template comprising a part of the wild-type C9ORF72 gene or minigene or cDNA.
The method of any one of the preceding claims, wherein the C9ORF72 gene is located on Chromosome 9:
- 27,546,542-27,573,863 (Genome Reference Consortium—GRCh38/hg38).
One or more guide ribonucleic acids (gRNAs) comprising a spacer sequence selected from the group consisting of the nucleic acid sequences in SEQ ID NOs:
- 1-18807 for editing the C9ORF72 gene in a cell from a patient with amyotrophic lateral sclerosis (ALS) or frontotemporal lobular dementia (FTLD).
A method for treating a patient with amyotrophic lateral sclerosis (ALS) or frontotemporal lobular dementia (FTLD) comprising transplanting the bone marrow from a donor to the patient.
One or more guide ribonucleic acids (gRNAs) comprising a spacer sequence selected from the group consisting of the nucleic acid sequences in SEQ ID NOs:
- 18810-73668 for editing the C9ORF72 gene in a cell from a patient with amyotrophic lateral sclerosis (ALS) or frontotemporal lobular dementia (FTLD).
Dependencies are inferred from the claim text.