Pluripotent cell lines and methods of use thereof

• US 8,669,048 B2
• Filed: 06/24/2009
• Issued: 03/11/2014
• Est. Priority Date: 06/24/2008
• Status: Active Grant

• Current Assignee: Parkinson's Institute, Board of Trustees of the Leland Stanford Junior University (Stanford Management Co.)
• Original Assignee: Parkinson's Institute
• Inventors: Reijo Pera, Renee Ann, Langston, J. William, Palmer, Theodore D., Schle, Birgitt
• Primary Examiner(s): Long, Scott
• Application Number:
  US12/459,019
• Publication Number:
  US 20100167286A1
• Time in Patent Office: 1,721 Days
• Field of Search: None
• US Class Current: 435/6
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2500/46 Amines, e.g. putrescine
• CPC Class Codes: C12N 2501/01 Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/119 Other fibroblast growth factors, e.g. FGF-4, FGF-8, FGF-10
• CPC Class Codes: C12N 2501/13 Nerve growth factor [NGF]; Brain-derived neurotrophic factor [BDNF]; Cilliary neurotrophic factor [CNTF]; Glial-derived neurotrophic factor [GDNF]; Neurotrophins [NT]; Neuregulins
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/392 Sexual steroids
• CPC Class Codes: C12N 2501/41 Hedgehog proteins; Cyclopamine (inhibitor)
• CPC Class Codes: C12N 2501/50 Cell markers; Cell surface determinants
• CPC Class Codes: C12N 2501/602 Sox-2
• CPC Class Codes: C12N 2501/603 Oct-3/4
• CPC Class Codes: C12N 2501/604 Klf-4
• CPC Class Codes: C12N 2501/70 Enzymes
• CPC Class Codes: C12N 2501/71 Oxidoreductases (EC 1.)
• CPC Class Codes: C12N 2503/02 Drug screening
• CPC Class Codes: C12N 2506/1307 from adult fibroblasts
• CPC Class Codes: C12N 2506/45 from artificially induced pluripotent stem cells
• CPC Class Codes: C12N 2740/15032 Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
• CPC Class Codes: C12N 5/0619 : Neurons
• CPC Class Codes: C12N 7/00 : Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00
• CPC Class Codes: C12Q 1/6883 : for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/136 : Screening for pharmacological compounds
• CPC Class Codes: G01N 2333/47 : Assays involving proteins of known structure or function as defined in the subgroups
• CPC Class Codes: G01N 33/5023 : on expression patterns
• CPC Class Codes: G01N 33/5058 : Neurological cells

Abstract

Methods of generating cell lines with a sequence variation or copy number variation of a gene of interest, methods of use thereof, and cell lines with a sequence variation or copy number variation of a gene of interest are provided.

Claims

1. A method of determining whether an agent is useful in the treatment of Parkinson'"'"'s disease or Parkinson'"'"'s-related disease, the method comprising:

a) contacting dopaminergic cells produced from induced pluripotent stem cells derived from a patient with Parkinson'"'"'s disease or Parkinson'"'"'s-like disease with an agent;

b) detecting a response to said agent in said cells; and

c) determining whether the agent is useful in the treatment based on the detected response.

2. The method of claim 1 wherein the agent is selected from a small molecule, a drug, an antibody, a hybrid antibody, an antibody fragment, a siRNA, an antisense RNA, an aptamer, a protein, or a peptide.

3. The method of claim 1 wherein the agent corrects an α

-synuclein dysfunction.

4. The method of claim 3 wherein the agent is selected from the group consisting of apomorphine, pyrogallol, 1,4-naphthoquinone, cisplatin, isoproterenol, pyrogallin, cianidanol, sulfasalazine, quinalizarin, benserazide, hexachlorophene, pyrvinium pamoate, dobutamine, methyl-dopa, curcumin, berberine chloride, daidzein, merbromin, norepinephrine, dopamine hydrochloride, carbidopa, ethylnorepinephrine hydrochloride, tannic acid, elaidyphosphocholine, hydroquinone, chlorophyllide Cu complex Na salt, methyldopa, isoproterenol hydrochloride, benserazide hydrochloride, dopamine, dobutamine hydrochloride, thyroid hormone, purpurin, sodium beta-nicotinamide adenine dinucleotide phosphate, lansoprazole, dyclonine hydrochloride, pramoxine hydrochloride, azobenzene, cefamandole sodium, cephaloridine, myricetin, 6,2′

,3′

-trihydroxyflavone, 5,7,3′

,4′

,5′

-pentahydroxyflavone, 7,3′

,4′

,5′

-tetrahydroxyflavone, (5,6,7,4′

-tetrahydroxyflavone), baicalein, eriodictyol, 7,3′

,4′

-trihydroxyisoflavone, epigallocatechin gallate, quercetin, gossypetin (3,5,7,8,3′

,4′

-hexahydroxyflavone), 2′

,3′

-dihydroxyflavone, 3′

,4′

-dihydroxyflavone, 5,6-dihydroxy-7-methoxyflavone, baicalein-7-methyl ether, 1-dopa, DOPAC, homogentisic acid, 6-hydroxydopamine, epinephrine, 3,4-dihydroxycinnamic acid, 2,3-dihydroxynaphthalene, 3,4-dihydroxybenzoic acid, 3,4,5-trihydroxybenzoic acid, 1,2,3-trihydroxybenzoic acid, gallate (gallic acid), benzoquinone, catechol, rifampicin, rosmarinic acid, baicalin, tanshinones I and II, emodin, procyanidin B4, resveratrol, rutin, fisetin, luteolin, fustin, epicatechin gallate, catechin, alizarin, tannic acid, eriodyctol, carboplatin, purpurogallin-4-carboxylic acid, koparin, 2,3,4-trihydroxy-4′

-ethexybenzophenone, baeomycesic acid, hamtoxylin, iriginol hexaaceatate, 4-acetoxyphenol, theaflavin monogallate, theaflavin digallate, stictic acid, purpurogallin, 2,5-dihydroxy-3,4-dimethoxy-4′

-ethoxybenzophenone, promethazine hydrochloride, oxidopamine hydrochloride, pyrantel pamoate, elaidylphosphocholine, amphotericin B, gallic acid, fumarprotocetraric acid, theaflavin, haematoxylin pentaacetate, 4-methoxydalbergione, epigallocatechin-3-monogallate, rolitetracycline, 7,3′

-dimethoxyflavone, liquiritigenin dimethyl ether, catechin pentaacetate, apigenin, 3,4-dedesmethyl-5-deshydroxy-3′

-ethoxyscleroin, derivatives and analogs thereof.

5. The method of claim 1 wherein the induced pluripotent stem cells are derived from human fibroblasts.

6. The method of claim 1 wherein the induced pluripotent stem cells are produced with a method comprising the use of three factors.

7. The method of claim 6 wherein the three factors are OCT4, SOX2, and KLF4.

8. The method of claim 1 wherein the patient carries a genetic variation or mutation known to be associated with Parkinson'"'"'s disease or Parkinson'"'"'s-like disease.

9. The method of claim 8, wherein the genetic variation of interest is a copy number variation of gene of interest.

10. The method of claim 9, wherein the cell line has three copies of the gene of interest.

11. The method of claim 10 wherein the gene of interest is PARK1 (SNCA or α

-synuclein).

12. The method of claim 8 wherein the gene of interest is PARK8 (LRRK2).

13. The method of claim 12 wherein the patient carries a homozygous mutation of LRRK2.

14. The method of claim 13 wherein the mutation comprises a G2019S mutation.

15. The method of claim 1 wherein the patient has Parkinson'"'"'s disease.

16. The method of claim 1, wherein said detected response is a change in cell viability, cellular chemistry, cellular function, mitochondrial function, cell aggregation, cell morphology, cellular protein aggregation, gene expression, cellular secretion, or cellular uptake.