DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Election/Restrictions
Applicant’s election without traverse of Group I (claims 1-11) in the reply filed on 12/08/2025 is acknowledged.
Claims 12-17 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 12/08/2025.
Claims 1-11 are under examination.
Priority
This application claims foreign priority to Japan 2022-046107, filed 03/22/2022.
Receipt is acknowledged of certified copies of papers required by 37 CFR 1.55. However, there is no English translation of the non-English language foreign priority application. Therefore, the instant application receives the priority date of the filing date, 03/20/2023.
Specification
The disclosure is objected to because it contains an embedded hyperlink and/or other form of browser-executable code. Applicant is required to delete the embedded hyperlink and/or other form of browser-executable code; references to websites should be limited to the top-level domain name without any prefix such as http:// or other browser-executable code. See MPEP § 608.01. See paragraph 0060, reciting “http://” and “https://”.
Claim Objections
Claims 8-10 are objected to because of the following informalities: Claims 8-10 each recite “MECP2”. At least the first recitation of the full name of the gene should be fully written out. Appropriate correction is required.
Claim Rejections - 35 USC § 112
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Written Description Rejection
Claims 1-11 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for pre-AIA the inventor(s), at the time the application was filed, had possession of the claimed invention.
Claims 1-3 encompass a method for producing any in vitro genetic disease model cell, comprising: inhibiting by any means, a genus of responsible genes of a genus of genetic diseases of interest from being expressed in any subject differentiated cell derived from any pluripotent stem cell or any adult stem cell. Claim 3 requires the inducing to be performed simultaneously with the inhibiting. Claim 4 recites expression of the responsible gene is inhibited using a gene knockdown method, however this still encompasses a genus of gene knockdown methods, including RNAi, antisense oligonucleotides, aptamers, miRNAs, CRISPR gene editing. Claim 5 limits the gene knockdown method to RNAi, antisense oligonucleotide and a nucleic acid aptamer method. Claim 6 limits the genetic disease of interest to a central nervous system disease and the subject differentiated cell to a neurocyte, however this claim still encompasses a large genus of central nervous system diseases. Claims 8-10 further limit the central nervous system disease to Rett syndrome and the responsible gene to MECP2 gene and recites specific amino acid and gene sequences. Claim 11 defines the pluripotent stem cell to an iPS cell or ES cell.
Regarding the state of the art of inhibiting gene expression, Tsai et al. (Journal of Translational Medicine, 2022, 20:535) teach that abnormal gene expression level or expression of genes containing deleterious mutations are two of the main determinants which lead to genetic disease, and to obtain a therapeutic effect and cure genetic diseases, it is crucial to regulate the host’s gene expression and restore it to physiological conditions (Abstract).
Tsai et al. teach there are a variety of molecular tools to regulate gene expression, including genome editing nucleases, transposons, episomes, siRNA, and shRNA (page 2, left column). Tsai et al. taught gene-editing nucleases include meganucleases (MNs), zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and CRISPR associated endonucleases, and to guide the nuclease to the target site, MNs, ZFNs and TALENs use a protein-DNA interaction and CRISPR-Cas systems are guided by RNA-DNA interactions (page 2, right column). Tsai et al. taught episomes are closed circular DNA molecules are autonomously replicating (page 6, right column).
Tsai et al. taught RNA interference is mediated by three classes of molecules: siRNA, shRNA and bifunctional shRNA, with siRNA being a double-stranded RNA molecules typically 20-24 base pairs in length, and after being delivered to the cell an incorporated into the RNA-Induced Silencing Complex (RISC) where it is unwound to a single strand RNA, and the less thermodynamically stable RNA strand is then used by RIS to probe and anneal with target complementary mRNA, which is subsequently cleaved (page 7, right column). Tsai et al. taught shRNAs contain a tight hairpin turn and are encoded in a DNA vector to be delivered to cells (page 7, right column).
Therefore, Tsai et al. teach different means of inhibiting target gene expression, which each have different chemical structures and mechanisms of action.
Ross et al. (Hum Mol Genet. 2014 May 13; 23(R1); R17-R26) teach cell biology of human neurodegenerative diseases have been difficult to study, but that development of human induced pluripotent stem cell models has greatly enhanced the ability to model disease in human cells, and that methods have been improved including increasing reprogramming efficiency, introducing non-viral and non-integrating methods of cell reprogramming, and using novel gene editing techniques for generating genetically corrected lines from patient-derived iPSCs, or for generating mutations in control cell lines (Abstract). Ross et al. teach different iPSC models can be used to study various neurodegenerative disorders such as Huntington's disease, Parkinson's disease, Amyotrophic Lateral Sclerosis, Fronto-Temporal Dementia, Alzheimer's disease, Spinomuscular Atrophy and other polyglutamine diseases (Abstract).
Ross et al. teach critical for the ability to model neurodegenerative diseases is the capacity to generate defined neuronal populations from pluripotent cells. While much progress has been made, current protocols are still often lengthy, expensive and complex and yield incompletely homogeneous populations of neurons. Nevertheless, it has become possible to differentiate human pluripotent stem cells into cells with phenotypes resembling dopaminergic neurons, glutamatergic neurons, GABAergic neurons, motor neurons and medium spiny neurons of the striatum, and human pluripotent stem cells can also be differentiated in vitro into astroglial and oligodendrocyte precursors able to further differentiation into mature astrocytes and oligodendrocytes in vivo (pages R17-R18).
Ross et al. teach the opportunities for studying genetic neurodegenerative diseases have also been greatly enhanced by modern techniques of genome editing. These include homologous recombination and the more recent and efficient techniques involving zinc finger nucleases, transcription activator-like effector nucleases (TALENs) and the highly efficient and low-cost technique termed Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR). The CRISPR/Cas9 system consists of the CRISPR-associated (Cas) nuclease Cas9 and a short guide RNA (gRNA) to target the bacterial double-stranded DNA endonuclease Cas9 to specific genomic sequence. These genome-editing techniques have many potential applications for the studying of neurodegenerative diseases, including correction or generation of genetic mutations, and addition of experimental alterations to cell lines with genetic mutations, with applications for disease modeling, or potentially for cell transplantation approaches (page R18).
Therefore, Ross et al. teach numerous diseases that are considered neurodegenerative diseases, that gene editing can be used to study genetic neurodegenerative diseases, and differentiation of pluripotent stem cells in vitro into astroglial and oligodendrocyte precursors.
The specification discloses production of a Rett Syndrome Model Cell comprising inhibiting the hMECP2 gene responsible for the genetic disease Rett Syndrome, using shRNA of SEQ ID NO: 3 in iPS-cell derived neurocytes and human astrocytes (Example 1, pages 28-30), which meets the written description and enablement provisions of 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph. However, claims 1-11 are directed to encompass a genus of means of inhibiting a genus of responsible genes of a genus of genetic diseases of interest from being expressed in any subject differentiated cell, which only correspond in some undefined way to specifically instantly disclosed genes, genetic diseases, and inhibitors and cells. With the exception of the above specifically disclosed gene (hMECP2), genetic disease of interest (Rett Syndrome), shRNA inhibitor of SEQ ID NO: 3, and the differentiated cells being neurocytes, the method comprising inhibiting a responsible gene of a genetic disease of interest from being expressed in a subject differentiated cell derived from a pluripotent stem cell or adult stem cell does meet the written description provision of 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, due to lacking chemical structural information for what they are and chemical structures are highly variant and encompass a myriad of possibilities. The specification provides insufficient written description to support the genus encompassed by the claim. Note: MPEP 2163.
Vas-Cath Inc. v. Mahurkar, 19 USPQ2d 1111, (Fed. Cir. 1991), makes clear that "applicant must convey with reasonable clarity to those skilled in the art that, as of the filing date sought, he or she was in possession of the invention. The invention is, for purposes of the 'written description' inquiry, whatever is now claimed." (See page 1117.) The specification does not "clearly allow persons of ordinary skill in the art to recognize that [he or she] invented what is claimed." (See Vas-Cath at page 1116.)
Univ. of Rochester v. G.D. Searle, 69 USPQ2d 1886, 1892 (CAFC 2004), further supports this by stating that:
The appearance of mere indistinct words in a specification or a claim, even an original claim, does not necessarily satisfy that requirement. A description of an anti-inflammatory steroid, i.e., a steroid (a generic structural term) described even in terms of its functioning of lessening inflammation of tissues fails to distinguish any steroid from others having the same activity or function. A description of what a material does, rather than of what it is, usually does not suffice…. The disclosure must allow one skilled in the art to visualize or recognize the identity of the subject matter purportedly described. (Emphasis added).
With the exception of the above specifically disclosed gene (hMECP2), genetic disease of interest (Rett Syndrome), shRNA inhibitor of SEQ ID NO: 3 and the subject differentiated cell being an iPS-cell derived neurocyte, the skilled artisan cannot envision the detailed chemical structure of the encompassed inhibitors that inhibit responsible genes of a genetic disease of interest or the genus of subject differentiated cells. Adequate written description requires more than a mere statement that it is part of the invention and reference to a potential method for isolating it. The chemical structure itself is required. See Fiers v. Revel, 25 USPQ2d 1601, 1606 (Fed. Circ. 1993) and Amgen Inc. V. Chugai Pharmaceutical Co. Ltd., 18 USPQ2d 1016, (Fed. Cir. 1991). In Fiddes v. Baird, 30 USPQ2d 1481, 1483, (Bd. Pat. App. & Int. 1993), claims directed to mammalian FGF's were found unpatentable due to lack of written description for the broad class. The specification provided only the bovine sequence. Finally, University of California v. Eli Lilly and Co., 43 USPQ2d 1398, 1404, 1405 (Fed. Cir. 1997) held that:
...To fulfill the written description requirement, a patent specification must describe an invention and do so in sufficient detail that one skilled in the art can clearly conclude that "the inventor invented the claimed invention." Lockwood v. American Airlines, Inc., 107 F.3d 1565, 1572, 41 USPQ2d 1961, 1966 (Fed. Cir. 1997); In re Gosteli, 872 F.2d 1008, 1012, 10 USPQ2d 1614, 1618 (Fed. Cir. 1989) (" [T]he description must clearly allow persons of ordinary skill in the art to recognize that [the inventor] invented what is claimed."). Thus, an applicant complies with the written description requirement "by describing the invention, with all its claimed limitations, not that which makes it obvious," and by using "such descriptive means as words, structures, figures, diagrams, formulas, etc., that set forth the claimed invention." Lockwood, 107 F.3d at 1572, 41 USPQ2d at 1966.
Furthermore, to the extent that a functional description can meet the requirement for an adequate written description, it can do so only in accordance with PTO guidelines stating that the requirement can be met by disclosing “sufficiently detailed, relevant identifying characteristics,” including “functional characteristics when coupled with a known or disclosed correlation between function and structure.” Univ. of Rochester v. G.D. Searle, 68 USPQ2d 1424, 1432 (DC WNY 2003). In this case, the specification does not provide a core structure of the inhibitor that inhibits a responsible gene of a genetic disease of interest from being expressed in a subject differentiated cells derived from a pluripotent stem cell or adult stem cell, and there is no structure-function correlation of the inhibitor that results in the claimed function.
Therefore, only the above chemically structurally defined chemicals, but not the full breadth of the claim(s) meet the written description provision of 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph. The species specifically disclosed are not representative of the genus because the genus is highly variant. Applicant is reminded that Vas-Cath makes clear that the written description provision of 35 USC § 112 is severable from its enablement provision. (See page 1115.)
Claim Rejections - 35 USC § 102
The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention.
Claims 1,2,4-6,8 and 11 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Marchetto et al. (WO 2011079307, Published 30 June 2011), cited on an IDS.
Regarding claims 1,4-6,8 and 11, Marchetto et al. taught a cellular approach to study autism spectrum disorders (ASD), using Rett syndrome (RTT) as an ASD genetic model, Marchetto et al. developed an in vitro system by deriving induced pluripotent stem cells from a RTT patient’s fibroblast (paragraph 0071). Marchetto et al. taught most RTT patients have mutations in the X-linked gene encoding MeCP2, and that RTT patient’s iPSCs are pluripotent and able to generate proliferating neural progenitor cells and functional postmitotic neurons and that an in vitro model of RTT was generated (paragraph 0071). Marchetto et al. taught shRNA against a target sequence of the human MeCP2 gene was cloned in a lentivirus vector, and
after infection, fibroblasts were plated with hESC medium (paragraph 0072). Marchetto et al. also taught testing whether a loss of function of MeCP2 was directly related to the number of glutamatergic synapses in the neuronal cultures, and cloned shRNA against MeCP2 in a lentiviral vector that is able to knockdown both isoforms of MeCP2, and neurons derived from WT-iPSCs expressing the shMeCP2 showed a reduction in VGLUT1 puncta. Therefore, Marchetto et al. taught inhibiting a responsible gene (MeCP2) of a genetic disease of interest (Rett syndrome) from being expressed by an RNAi method using shRNA against MeCP2 in neurons derived from iPSCs.
Regarding claim 2, Marchetto et al. taught iPSC colonies were directly transferred to feeder-free conditions on Matrigel-coated dishes; to obtain neural progenitor cells (NPCs), embryoid bodies were formed by mechanical dissociation of cell clusters and plating onto low-adherence dishes in hESC medium, then plated onto poly-ornithine/laminin coated dishes….and populations of NPCs were achieved. To obtain mature neurons, floating EBs were treated with retinoic acid (paragraph 0072).
Regarding claim 7, Marchetto et al. taught neurons co-cultured with astrocytes (a glia cell) (paragraph 0081), and that astrocytes were present in the cultures with the neurons (paragraph 0101). The instant specification discloses that examples of known glia cells include an astrocyte, Schwann cell, oligodendrocyte and satellite cell (paragraph 0026).
Claims 1,2,4-8 and 11 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Ohashi (UCLA Electronic Theses and Dissertations, Published 2017), cited on an IDS.
Regarding claims 1,2,4-8 and 11 Ohashi taught that induced pluripotent stem cells have created new possibilities to more accurately model human disease in a dish, and that iPSCs can both self-renew indefinitely and can differentiate into almost any cell lineage of three primary germ layers, which enables programming of diseased patient’s own fibroblasts into a pluripotent state, and then re-differentiating the cells into disease relevant cell types (page 11). Ohashi taught using iPSC models to study Rett syndrome, and the iPSC lines either expressed MECP2 or lacked this protein (page 12, first paragraph, page 39, first paragraph). Ohashi taught generating neural progenitor cells from the hiPSCs lines, which were then further differentiated to yield both neurons and glia (pages 41-42,58). Ohashi taught silencing MECP2 in WT NPCs, in which several different siRNA targeting oligos were assayed for their ability to silence MECP2, and that silencing of MECP2 in WT-NPCs led to strong induction of expression of subtelomeric genes (Pages 72-73).
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
Claim 3 is rejected under 35 U.S.C. 103 as being unpatentable over Marchetto et al. as applied to claims 1,2,4-6,8 and 11 above, and further in view of
Ross et al. (Hum Mol Genet. 2014 May 13; 23(R1); R17-R26).
The teachings of Marchetto et al. as applicable to claims 1,2,4-6,8 and 11 have been described above.
Marchetto et al. do not teach wherein inducing is performed simultaneously with the inhibiting.
Before the effective filing date, Ross et al. taught the cell biology of human neurodegenerative diseases have been difficult to study, but that development of human induced pluripotent stem cell models has greatly enhanced the ability to model disease in human cells, and that methods have been improved including increasing reprogramming efficiency, introducing non-viral and non-integrating methods of cell reprogramming, and using novel gene editing techniques for generating genetically corrected lines from patient-derived iPSCs, or for generating mutations in control cell lines (Abstract). Ross et al. taught different iPSC models can be used to study various neurodegenerative disorders such as Huntington's disease, Parkinson's disease, Amyotrophic Lateral Sclerosis, Fronto-Temporal Dementia, Alzheimer's disease, Spinomuscular Atrophy and other polyglutamine diseases (Abstract). Ross et al. taught critical for the ability to model neurodegenerative diseases is the capacity to generate defined neuronal populations from pluripotent cells. While much progress has been made, current protocols are still often lengthy, expensive and complex and yield incompletely homogeneous populations of neurons. Nevertheless, it has become possible to differentiate human pluripotent stem cells into cells with phenotypes resembling dopaminergic neurons, glutamatergic neurons, GABAergic neurons, motor neurons and medium spiny neurons of the striatum, and human pluripotent stem cells can also be differentiated in vitro into astroglial and oligodendrocyte precursors able to further differentiation into mature astrocytes and oligodendrocytes in vivo (pages R17-R18).
Ross et al. taught the opportunities for studying genetic neurodegenerative diseases have also been greatly enhanced by modern techniques of genome editing, including homologous recombination and the more recent and efficient techniques involving zinc finger nucleases, transcription activator-like effector nucleases (TALENs) and the highly efficient and low-cost technique termed Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR). Ross et al. taught these genome-editing techniques have many potential applications for the studying of neurodegenerative diseases, including correction or generation of genetic mutations, and addition of experimental alterations to cell lines with genetic mutations, with applications for disease modeling, or potentially for cell transplantation approaches (page R18).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date, to have modified the in vitro cell model production method of Marchetto et al. to make integral the step of inducing the pluripotent stem cells into the subject differentiated cell and inhibiting the disease gene based on the teachings of Ross et al. regarding the problems with current protocols still being often lengthy, expensive and complex. There would be a reasonable expectation of success, because both Marchetto et al. and Ross et al. pertain to producing induced pluripotent stem cells as in vitro models of neurodegenerative disease. One of ordinary skill in the art would have been motivated to modify the in vitro cell model production method of Marchetto et al. to make integral the step of inducing the pluripotent stem cells into the subject differentiated cell and inhibiting the disease gene (the inducing is performed simultaneously with the inhibiting), rather than occurring in different steps, because Ross et al. taught iPSC models can be used to study various neurodegenerative disorders but current protocols are still often lengthy, expensive and complex. Therefore, it would have been obvious to combine and make integral the inducing differentiation and inhibiting into the same step, rather than in multiple steps, to reduce time in creating the in vitro cell model, which would reduce expenses and create a less complex production method, which is taught as a problem to be solved in the prior art. See MPEP 2144.04 V. B.
Therefore, the invention as a whole would have been prima facie obvious to one of ordinary skill in the art before the effective filing date.
Claims 9 and 10 are rejected under 35 U.S.C. 103 as being unpatentable over Marchetto et al. as applied to claims 1,2,4-6,8 and 11 above, and further in view of Minassian et al. (US 20060194257, Published 31 Aug 2006).
The teachings of Marchetto et al. as applicable to claims 1,2,4-6,8 and 11 have been described above.
Marchetto et al. do not teach the amino acid sequence or nucleotide sequence of MeCP2.
Before the effective filing date, Minassian et al. taught that up to 80% of patients with Rett syndrome have mutations in exons 3 and 4 of the 4-exon MECP2 gene, and the need for identification of further mutations to account for the remaining 20% of RTT patients so methods of diagnosing and treating RTT can be identified (paragraphs 0004 and 0006). Minassian et al. taught the identification of a novel open reading frame of the MECP2 gene, called MECP2E1. Minassian et al. taught that patients with a neuropsychiatric disorder or developmental disorder such as Rett’s syndrome and mental retardation had mutations in exon 1 of the MECP2E1 gene and provides methods of detecting the disorder by detecting the mutation or deletion in exon 1 of the MECP2E1 sequence (SEQ ID NO: 3) (paragraph 0018).
Minassian et al. taught an isolated MeCP2E1 protein encoded by the nucleic acid molecules of the invention, and that the MeCP2E1 protein has the amino acid sequence as shown in Fig. 6(b), SEQ ID NO: 4 (paragraph 0065).
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The amino acid sequence of SEQ ID NO: 4 of the MeCP2E1 protein of Minassian et al. has 100% identity to instant SEQ ID NO: 1. Below is the alignment between the amino acid sequence of instant SEQ ID NO: 1 (Qy) and the amino acid sequence (SEQ ID NO: 4) of Minassian et al. (Db):
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Minassian et al. taught an isolated nucleic acid molecule comprising a sequence encoding the MECP2E1 protein, and that the purified and isolated nucleic acid molecule comprises a nucleic acid sequence encoding a protein as shown in Fig. 6(b), SEQ ID NO: 4 (paragraphs 0010-0011), and that the isolated nucleic acid molecule has a sequence as shown in Fig. 6(a) (SEQ ID NO: 3 or a fragment or variant thereof (paragraphs 0041,0042).
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The nucleic acid sequence of SEQ ID NO: 3 of Minassian et al. has 100% identity to instant SEQ ID NO: 2. Below is the alignment between the sequence of instant SEQ ID NO: 2 (Qy) and the SEQ ID NO: 3 of Minassian et al. (Db):
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Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention, that the MeCP2 amino acid sequence and nucleotide sequence pertaining to the in vitro cell model production method of Marchetto et al., would have had the amino acid sequence of SEQ ID NO: 4 and the gene would have the sequence of SEQ ID NO: 3 of Minassian et al. with a reasonable expectation of success. There would be a reasonable expectation of success, because both Marchetto et al. and Minassian et al. pertain to Rett syndrome caused by mutations in the X-linked gene encoding MeCP2 protein, and would amount to simple substitution of the MeCP2 protein and gene of Marchetto et al. with the known MeCP2 amino acid and gene sequences of Minassian et al. to obtain predictable results. One of ordinary skill in the art would have been motivated to have used the amino acid of SEQ ID NO: 4 and the nucleotide sequence of SEQ ID NO: 3 of Minassian et al. as the sequences in the method of Marchetto et al., because Minassian et al. taught the need for identification of further mutations in MECP2 to account for the remaining 20% of RTT patients so methods of diagnosing and treating RTT can be identified, and Minassian et al. taught identification of a novel open reading frame of the MECP2 gene, called MECP2E1 comprising the amino acid sequence of SEQ ID NO: 4 and the nucleotide sequence of SEQ ID NO: 3. Minassian et al. taught that patients with a neuropsychiatric disorder or developmental disorder such as Rett’s syndrome and mental retardation had mutations in exon 1 of the MECP2E1 gene and provides methods of detecting the disorder by detecting the mutation or deletion in exon 1 of the MECP2E1 sequence (SEQ ID NO: 3) (paragraph 0018). Therefore, SEQ ID NOs: 3 and 4 of Minassian et al. would have been suitable sequences to use in the method of Marchetto et al.
Accordingly, the limitations of claims 9 and 10 would have been prima facie obvious to one of ordinary skill in the art before the effective filing date.
Claim 3 is rejected under 35 U.S.C. 103 as being unpatentable over Ohashi as applied to claims 1,2,4-8 and 11 above, and further in view of
Ross et al. (Hum Mol Genet. 2014 May 13; 23(R1); R17-R26).
The teachings of Ohashi as applicable to claims 1,2,4-8 and 11 have been described above.
Ohashi does not teach wherein inducing is performed simultaneously with the inhibiting.
Before the effective filing date, Ross et al. taught the cell biology of human neurodegenerative diseases have been difficult to study, but that development of human induced pluripotent stem cell models has greatly enhanced the ability to model disease in human cells, and that methods have been improved including increasing reprogramming efficiency, introducing non-viral and non-integrating methods of cell reprogramming, and using novel gene editing techniques for generating genetically corrected lines from patient-derived iPSCs, or for generating mutations in control cell lines (Abstract). Ross et al. taught different iPSC models can be used to study various neurodegenerative disorders such as Huntington's disease, Parkinson's disease, Amyotrophic Lateral Sclerosis, Fronto-Temporal Dementia, Alzheimer's disease, Spinomuscular Atrophy and other polyglutamine diseases (Abstract). Ross et al. taught critical for the ability to model neurodegenerative diseases is the capacity to generate defined neuronal populations from pluripotent cells. While much progress has been made, current protocols are still often lengthy, expensive and complex and yield incompletely homogeneous populations of neurons. Nevertheless, it has become possible to differentiate human pluripotent stem cells into cells with phenotypes resembling dopaminergic neurons, glutamatergic neurons, GABAergic neurons, motor neurons and medium spiny neurons of the striatum, and human pluripotent stem cells can also be differentiated in vitro into astroglial and oligodendrocyte precursors able to further differentiation into mature astrocytes and oligodendrocytes in vivo (pages R17-R18).
Ross et al. taught the opportunities for studying genetic neurodegenerative diseases have also been greatly enhanced by modern techniques of genome editing, including homologous recombination and the more recent and efficient techniques involving zinc finger nucleases, transcription activator-like effector nucleases (TALENs) and the highly efficient and low-cost technique termed Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR). Ross et al. taught these genome-editing techniques have many potential applications for the studying of neurodegenerative diseases, including correction or generation of genetic mutations, and addition of experimental alterations to cell lines with genetic mutations, with applications for disease modeling, or potentially for cell transplantation approaches (page R18).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date, to have modified the in vitro cell model production method of Ohashi to make integral the step of inducing the pluripotent stem cells into the subject differentiated cell and inhibiting the disease gene based on the teachings of Ross et al. regarding the problems of current protocols still being often lengthy, expensive and complex. There would be a reasonable expectation of success, because both Ohashi and Ross et al. pertain to producing induced pluripotent stem cells as in vitro models of neurodegenerative disease. One of ordinary skill in the art would have been motivated to modify the in vitro cell model production method of Ohashi to make integral the step of inducing the pluripotent stem cells into the subject differentiated cell and inhibiting the disease gene (the inducing is performed simultaneously with the inhibiting), rather than occurring in different steps, because Ross et al. taught iPSC models can be used to study various neurodegenerative disorders but current protocols are still often lengthy, expensive and complex. Therefore, it would have been obvious to combine and make integral the inducing differentiation and inhibiting into the same step, rather than in multiple steps, to reduce time in creating the in vitro cell model, which would reduce expenses and create a less complex production method, which is taught as a problem to be solved in the prior art. See MPEP 2144.04 V. B.
Therefore, the invention as a whole would have been prima facie obvious to one of ordinary skill in the art before the effective filing date.
Claims 9 and 10 are rejected under 35 U.S.C. 103 as being unpatentable over Ohashi as applied to claims 1,2,4-8 and 11 above, and further in view of Minassian et al. (US 20060194257, Published 31 Aug 2006).
The teachings of Ohashi as applicable to claims 1,2,4-8 and 11 have been described above.
Ohashi does not teach the amino acid sequence or nucleotide sequence of MeCP2.
Before the effective filing date, Minassian et al. taught that up to 80% of patients with Rett syndrome have mutations in exons 3 and 4 of the 4-exon MECP2 gene, and the need for identification of further mutations to account for the remaining 20% of RTT patients so methods of diagnosing and treating RTT can be identified (paragraphs 0004 and 0006). Minassian et al. taught the identification of a novel open reading frame of the MECP2 gene, called MECP2E1. Minassian et al. taught that patients with a neuropsychiatric disorder or developmental disorder such as Rett’s syndrome and mental retardation had mutations in exon 1 of the MECP2E1 gene and provides methods of detecting the disorder by detecting the mutation or deletion in exon 1 of the MECP2E1 sequence (SEQ ID NO: 3) (paragraph 0018).
Minassian et al. taught an isolated MeCP2E1 protein encoded by the nucleic acid molecules of the invention, and that the MeCP2E1 protein has the amino acid sequence as shown in Fig. 6(b), SEQ ID NO: 4 (paragraph 0065).
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The amino acid sequence of SEQ ID NO: 4 of the MeCP2E1 protein of Minassian et al. has 100% identity to instant SEQ ID NO: 1. Below is the alignment between the amino acid sequence of instant SEQ ID NO: 1 (Qy) and the amino acid sequence (SEQ ID NO: 4) of Minassian et al. (Db):
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Minassian et al. taught an isolated nucleic acid molecule comprising a sequence encoding the MECP2E1 protein, and that the purified and isolated nucleic acid molecule comprises a nucleic acid sequence encoding a protein as shown in Fig. 6(b), SEQ ID NO: 4 (paragraphs 0010-0011), and that the isolated nucleic acid molecule has a sequence as shown in Fig. 6(a) (SEQ ID NO: 3 or a fragment or variant thereof (paragraphs 0041,0042).
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527
734
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The nucleic acid sequence of SEQ ID NO: 3 of Minassian et al. has 100% identity to instant SEQ ID NO: 2. Below is the alignment between the sequence of instant SEQ ID NO: 2 (Qy) and the SEQ ID NO: 3 of Minassian et al. (Db):
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431
816
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273
815
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Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention, that the MeCP2 amino acid sequence and nucleotide sequence pertaining to the in vitro cell model produced by Ohashi would have had the amino acid sequence of SEQ ID NO: 4 and the gene would have the sequence of SEQ ID NO: 3 of Minassian et al. with a reasonable expectation of success. There would be a reasonable expectation of success, because both Ohashi and Minassian et al. pertain to Rett syndrome caused by mutations in the X-linked gene encoding MeCP2 protein, and would amount to simple substitution of the MeCP2 protein and gene of Ohashi with the known MeCP2 amino acid and gene sequences of Minassian et al. to obtain predictable results. One of ordinary skill in the art would have been motivated to have used the amino acid of SEQ ID NO: 4 and the nucleotide sequence of SEQ ID NO: 3 of Minassian et al. as the sequences in the method of Ohashi because Minassian et al. taught the need for identification of further mutations in MECP2 to account for the remaining 20% of RTT patients so methods of diagnosing and treating RTT can be identified, and Minassian et al. taught identification of a novel open reading frame of the MECP2 gene, called MECP2E1 comprising the amino acid sequence of SEQ ID NO: 4 and the nucleotide sequence of SEQ ID NO: 3. Minassian et al. taught that patients with a neuropsychiatric disorder or developmental disorder such as Rett’s syndrome and mental retardation had mutations in exon 1 of the MECP2E1 gene and provides methods of detecting the disorder by detecting the mutation or deletion in exon 1 of the MECP2E1 sequence (SEQ ID NO: 3) (paragraph 0018). Therefore, SEQ ID NOs: 3 and 4 of Minassian et al. would have been suitable sequences to use in the method of Ohashi.
Accordingly, the limitations of claims 9 and 10 would have been prima facie obvious to one of ordinary skill in the art before the effective filing date.
Conclusion
Claims 1-11 are rejected.
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/STEPHANIE L SULLIVAN/Examiner, Art Unit 1635
/ABIGAIL VANHORN/Primary Examiner, Art Unit 1636