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 .
Response to Amendment/Status of Claims
Claims 30,31 and 49-52 were canceled. Claims 8,10,17 and 26 were amended. Claims 53-58 are new. Claims 1-3,5,7,8,10,15-17,23-26,29 and 53-58 are pending.
Election/Restrictions
Applicant's election with traverse of Group I (claims 1-3,5,7,8,10,15-17 and 53-56) in the reply filed on 07/29/2026 is acknowledged. While Applicant did not state with or without traverse, Applicant stated that in MPEP 803 if search and examination of an entire application can be made without serious burden, the examiner must examine the entire application on the merits, even though the entire applications includes claims to independent or distinct inventions, and it is Applicant’s positions that it would not be unduly burdensome to perform a search on all of the claims together in the present application. This is not found persuasive because MPEP 803 pertains to restriction for Applications filed under 35 U.S.C. 111 and this application is a 371. The Requirement of Unity of Invention is used in a 371 application and burden is not a consideration.
The requirement is still deemed proper and is therefore made FINAL.
Applicant’s election of SEQ ID NO: 32 for Species A as the amino acid sequence of the retrograde-enhanced clade E variant AAV capsid protein in the reply filed on 07/29/2026 is acknowledged. Because applicant did not distinctly and specifically point out the supposed errors in the restriction requirement, the election has been treated as an election without traverse (MPEP § 818.01(a)).
Upon further consideration, the elected sequence SEQ ID NO: 32 is free of the prior art, and therefore the species have been expanded to include SEQ ID NO: 33.
Claims 23-26,29,57 and 58 are withdrawn from further consideration pursuant to 37 CFR 1.142(b), as being drawn to a nonelected invention or species, there being no allowable generic or linking claim. Applicant timely traversed the restriction (election) requirement in the reply filed on 07/29/2026.
Claims 1-3,5,7,8,10,15-17 and 53-56 are under examination.
Priority
This application is a 371 of PCT/US2022/073560, filed 07/08/2022 which claims benefit of 63/221,759, filed 07/14/2021 as reflected by the most recent filing receipt.
Nucleotide and/or Amino Acid Sequence Disclosures
REQUIREMENTS FOR PATENT APPLICATIONS CONTAINING NUCLEOTIDE AND/OR AMINO ACID SEQUENCE DISCLOSURES
Items 1) and 2) provide general guidance related to requirements for sequence disclosures.
37 CFR 1.821(c) requires that patent applications which contain disclosures of nucleotide and/or amino acid sequences that fall within the definitions of 37 CFR 1.821(a) must contain a "Sequence Listing," as a separate part of the disclosure, which presents the nucleotide and/or amino acid sequences and associated information using the symbols and format in accordance with the requirements of 37 CFR 1.821 - 1.825. This "Sequence Listing" part of the disclosure may be submitted:
In accordance with 37 CFR 1.821(c)(1) via the USPTO patent electronic filing system (see Section I.1 of the Legal Framework for Patent Electronic System (https://www.uspto.gov/PatentLegalFramework), hereinafter "Legal Framework") as an ASCII text file, together with an incorporation-by-reference of the material in the ASCII text file in a separate paragraph of the specification as required by 37 CFR 1.823(b)(1) identifying:
the name of the ASCII text file;
ii) the date of creation; and
iii) the size of the ASCII text file in bytes;
In accordance with 37 CFR 1.821(c)(1) on read-only optical disc(s) as permitted by 37 CFR 1.52(e)(1)(ii), labeled according to 37 CFR 1.52(e)(5), with an incorporation-by-reference of the material in the ASCII text file according to 37 CFR 1.52(e)(8) and 37 CFR 1.823(b)(1) in a separate paragraph of the specification identifying:
the name of the ASCII text file;
the date of creation; and
the size of the ASCII text file in bytes;
In accordance with 37 CFR 1.821(c)(2) via the USPTO patent electronic filing system as a PDF file (not recommended); or
In accordance with 37 CFR 1.821(c)(3) on physical sheets of paper (not recommended).
When a “Sequence Listing” has been submitted as a PDF file as in 1(c) above (37 CFR 1.821(c)(2)) or on physical sheets of paper as in 1(d) above (37 CFR 1.821(c)(3)), 37 CFR 1.821(e)(1) requires a computer readable form (CRF) of the “Sequence Listing” in accordance with the requirements of 37 CFR 1.824.
If the "Sequence Listing" required by 37 CFR 1.821(c) is filed via the USPTO patent electronic filing system as a PDF, then 37 CFR 1.821(e)(1)(ii) or 1.821(e)(2)(ii) requires submission of a statement that the "Sequence Listing" content of the PDF copy and the CRF copy (the ASCII text file copy) are identical.
If the "Sequence Listing" required by 37 CFR 1.821(c) is filed on paper or read-only optical disc, then 37 CFR 1.821(e)(1)(ii) or 1.821(e)(2)(ii) requires submission of a statement that the "Sequence Listing" content of the paper or read-only optical disc copy and the CRF are identical.
Specific deficiencies and the required response to this Office Action are as follows:
Specific deficiency – Nucleotide and/or amino acid sequences appearing in the drawings are not identified by sequence identifiers in accordance with 37 CFR 1.821(d). Sequence identifiers for nucleotide and/or amino acid sequences must appear either in the drawings or in the Brief Description of the Drawings. See at least Fig. 7b, Fig. 12 with sequence not identified by a sequence identifier.
Required response – Applicant must provide:
Replacement and annotated drawings in accordance with 37 CFR 1.121(d) inserting the required sequence identifiers;
AND/OR
A substitute specification in compliance with 37 CFR 1.52, 1.121(b)(3) and 1.125 inserting the required sequence identifiers into the Brief Description of the Drawings, consisting of:
A copy of the previously-submitted specification, with deletions shown with strikethrough or brackets and insertions shown with underlining (marked-up version);
A copy of the amended specification without markings (clean version); and
A statement that the substitute specification contains no new matter.
Specific deficiency – Nucleotide and/or amino acid sequences appearing in the specification are not identified by sequence identifiers in accordance with 37 CFR 1.821(d). See at least page 71, paragraph 00227, where “LADQDYTKTA” and page 81 with primer sequences not identified by sequence identifiers. Applicant should ensure all sequences are identified by the correct sequence identifier.
Required response – Applicant must provide:
A substitute specification in compliance with 37 CFR 1.52, 1.121(b)(3) and 1.125 inserting the required sequence identifiers, consisting of:
A copy of the previously-submitted specification, with deletions shown with strikethrough or brackets and insertions shown with underlining (marked-up version);
A copy of the amended specification without markings (clean version); and
A statement that the substitute specification contains no new matter.
Drawings
The drawings are objected to because figure(s) 1-8,14 and 15 uses lower case letters to identify the different views of the figure whereas capital letters are required. See 37 CFR 1.84(u). (u) Numbering of views.
(1) The different views must be numbered in consecutive Arabic numerals, starting with 1, independent of the numbering of the sheets and, if possible, in the order in which they appear on the drawing sheet(s). Partial views intended to form one complete view, on one or several sheets, must be identified by the same number followed by a capital letter. View numbers must be preceded by the abbreviation "FIG.". For example, the correct label is FIG. 1A, FIG. 1B.
In addition, FIG. 10A shows the sequence of AAV8-retro as SEQ ID NO: 1. However, according to the Sequence Listing, SEQ ID NO: 1 is “LADQDYTKTA”. Applicant should provide the correct SEQ ID NO for AAV8-retro in FIG. 10A.
Corrected drawing sheets in compliance with 37 CFR 1.121(d) are required in reply to the Office action to avoid abandonment of the application. Any amended replacement drawing sheet should include all of the figures appearing on the immediate prior version of the sheet, even if only one figure is being amended. The figure or figure number of an amended drawing should not be labeled as “amended.” If a drawing figure is to be canceled, the appropriate figure must be removed from the replacement sheet, and where necessary, the remaining figures must be renumbered and appropriate changes made to the brief description of the several views of the drawings for consistency. Additional replacement sheets may be necessary to show the renumbering of the remaining figures. Each drawing sheet submitted after the filing date of an application must be labeled in the top margin as either “Replacement Sheet” or “New Sheet” pursuant to 37 CFR 1.121(d). If the changes are not accepted by the examiner, the applicant will be notified and informed of any required corrective action in the next Office action. The objection to the drawings will not be held in abeyance.
Specification
The disclosure is objected to because of the following informalities: There are many instances in the instant specification wherein the sequence “LADQDYTKTA” is identified as instant SEQ ID NO: 30. However, “QQQD” is also identified as SEQ ID NO: 30. Per the Sequence listing, “LADQDYTKTA” is SEQ ID NO: 1. See for Example page 2, paragraph 0010, page 3, paragraph 0012, page 21, paragraphs 0064-0065, pages 23-24, paragraph 0071, and pages 66-69 which incorrectly identifies “LADQDYTKTA” as instant SEQ ID NO: 30. There may be additional locations and the Examiner is only noting several locations and Applicant should ensure all instances are corrected.
Appropriate correction is required.
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 page 80 reciting “https://”.
Claim Objections
Claims 10 and 17 are objected to because of the following informalities: Claims 10 and 17 each recite “CamKv”. For clarity, the full name of the gene should be recited rather than just the acronym. Appropriate correction is required.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 1-3,5,7,8,10,15-17 and 53-56 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claims 1,2 and 5 recite the amino acid sequence “LADQDYTKTY (SEQ ID NO: 30)”. Claim 2 also recites an additional sequence “QQQD” which is also identified as SEQ ID NO: 30. This is indefinite and confusing, as both sequences cannot have the same SEQ ID NO while having different sequences. The Examiner noted this issue in the Restriction Requirement on page 4 that there was an error in the claims regarding this, and Applicant did not correct the issue in the new set of claims filed 07/29/2026. In addition, the specification also includes the same errors, so one looking to the specification would not know which sequence belongs to which SEQ ID NO: 1. Per the Sequence Listing, “LADQDYTKTY” is identified as SEQ ID NO: 1, and “QQQD” is identified as SEQ ID NO: 30. Therefore, in the interest of compact prosecution, the Examiner is interpreting “LADQDYTKTY” to be SEQ ID NO: 1 and not SEQ ID NO: 30, and “QQQD” as SEQ ID NO: 30.
Claims 3,7,8,10,15-17 and 53-56 depend on the rejected claims and do not correct the issue and are therefore included in the rejection.
The following is a quotation of 35 U.S.C. 112(d):
(d) REFERENCE IN DEPENDENT FORMS.—Subject to subsection (e), a claim in dependent form shall contain a reference to a claim previously set forth and then specify a further limitation of the subject matter claimed. A claim in dependent form shall be construed to incorporate by reference all the limitations of the claim to which it refers.
The following is a quotation of pre-AIA 35 U.S.C. 112, fourth paragraph:
Subject to the following paragraph [i.e., the fifth paragraph of pre-AIA 35 U.S.C. 112], a claim in dependent form shall contain a reference to a claim previously set forth and then specify a further limitation of the subject matter claimed. A claim in dependent form shall be construed to incorporate by reference all the limitations of the claim to which it refers.
Claims 7 and 10 are rejected under 35 U.S.C. 112(d) or pre-AIA 35 U.S.C. 112, 4th paragraph, as being of improper dependent form for failing to further limit the subject matter of the claim upon which it depends, or for failing to include all the limitations of the claim upon which it depends.
Claim 7 depends from claim 1 which recites a protein, with claim 7 reciting a nucleic acid encoding the protein. However, claim 7 does not further limit the protein itself. The Examiner suggests either amending claim 1 to recite the protein or a nucleic acid encoding the protein, or make claim 7 an independent claim.
Regarding claim 10, claim 10 depends on a later recited claim, 54, rather than a claim previously set forth.
Applicant may cancel the claim(s), amend the claim(s) to place the claim(s) in proper dependent form, rewrite the claim(s) in independent form, or present a sufficient showing that the dependent claim(s) complies with the statutory requirements.
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.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claims 1,5,7,15,16 and 56 are rejected under 35 U.S.C. 103 as being unpatentable over North Carolina (US 20190367562) in view of Hwang (WO 2017218842), both cited on an IDS.
Claim Interpretation: It is noted that there appears to be an error in the claims regarding the amino acid sequence “LADQDYTKTA” which is identified as SEQ ID NO: 30 in claim 1,2 and 5. However, according to the Sequence Listing, “LADQDYTKTA” is SEQ ID NO: 1. In the interest of compact prosecution, the Examiner is interpreting “LADQDYTKTA” to be SEQ ID NO: 1 rather than SEQ ID NO: 30 as claimed, and “QQQD” as SEQ ID NO: 30 per the sequence listing.
Regarding claims 1 and 5, North Carolina teaches modified adeno-associated virus capsid proteins comprising mutations (i.e., a modification, which can be a substitution, insertion or a deletion) in the amino acid sequence of a capsid protein from AAV8 or AAVrh.10 (Clade E) and teaches virus vectors comprising the modified AAV capsid protein (paragraphs 0004, 0079,0080). North Carolina teaches modified capsid proteins of the disclosure can be produced by modifying the capsid protein of any AAV including AAV8 and AAVrh.10 and those skilled in the art will understand that a variety of manipulations to the AAV capsid proteins are known in the art and the disclosure is not limited to modifications of naturally occurring AAV capsid proteins (paragraph 0112).
North Carolina teaches virus vectors used for retrograde transport (paragraph 0251), and with regard to the CNS, different AAV serotypes display a spectrum of transduction profiles and cellular tropisms, and that AAV capsids undergo axonal transport and transynaptic spread in the anterograde and/or retrograde directions depending on the serotype (paragraph 0254). North Carolina teaches a library of variant capsid genes using two serotypes, AAV1, and AAVrh.10, which is known to efficiently cross the BBB (paragraphs 0257,0274), and that AAVrh.10 falls within Clade E (paragraph 0308).
North Carolina does not teach a clade E variant AAV capsid protein comprising the amino acid sequence LADQDYTKTA.
Before the effective filing date, Hwang taught AAV variants that exhibit a preference for retrograde movement in neurons (Abstract, page 2, lines 28-30) and consensus sequences were identified that when present in an AAV capsid protein result in a profound enhancement in the efficacy of retrograde transport, and the consensus sequences are xxDxTKx (SEQ ID NO: 1) or xDxTKxx (SEQ ID NO: 2) (page 9, lines 21-23). Hwang taught while the capsid polypeptide of a virus particle can contain a consensus sequence as described herein, the remaining components can be from one or more known AAV serotypes (e.g. AAV2, AAV8, etc.) (page 13, lines 3-5). Hwang taught that four previously generated viral libraries were used at the start of the directed evolution procedure, including a DNA shuffling library generated from wild-type AAV8 cap gene sequences (page 17, Example 2). Hwang taught the specific peptide insert sequence for variants, including LADQDYTKTA for Clone 4R-SD-15 of SEQ ID NO: 32, for Clone 5R-Hind6 of SEQ ID NO: 44 (Table 2, page 20), for Clone 5R-Hind10 of SEQ ID NO: 48 (Table 2, page 21), Clone 5R-Hind17 of SEQ ID NO: 60 (Table 2, page 21), Clone 5R-Ce6 of SEQ ID NO: 72 (Table 2, page 21) shown below.
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Regarding claim 7, North Carolina teaches nucleic acid molecule encoding the modified virus capsids and capsid proteins of the disclosure (paragraph 0125).
Regarding claims 15,16 and 56, North Carolina teaches the terms “virus vector,” “vector” or “gene delivery vector” refer to a virus (e.g., AAV) particle that functions as a nucleic acid delivery vehicle, and which comprises the vector genome (e.g., viral DNA [vDNA]) packaged within a virion (paragraph 0068). North Carolina teaches AAV1/rh.10 chimeras capable of crossing the blood-brain barrier following intravenous administration and scans of immunostained brain sections showing GFP expression in the cerebral cortex mediated by parental or chimeric vectors packaging a CBh-scGFP transgene.
North Carolina does not explicitly teach a retrograde enhanced recombinant AAV particle comprising the retrograde-enhanced clade E variant AAV capsid protein of claim 1 and a nucleic acid comprising a transgene sequence.
The teachings of Hwang as applicable to claim 1 have been described above. In addition, Hwang taught a viral particle is provided that includes a viral capsid protein as described herein and that the viral particle exhibits a preference for retrograde movement and the viral particle possesses retrograde transport capability. A viral particle as described herein further includes a nucleic acid encoding a payload (the transgene sequence of instant claim 15) and that the coding sequence encoding the payload is selected from the group consisting of a protein-coding gene and an inhibitory RNA nucleic acid (the non-coding RNA or polypeptide of instant claim 16) (page 3, lines 18-28). Regarding claim 56, Hwang et al. taught the payload is an effector protein, including a gene editing system (e.g., CRISPR/Cas9, TALEN, zinc finger nuclease) (page 3, lines 30-31, page 4, line 1) and exemplifies SaCas9 in AAV2-retro (Example 12, page 26) and therefore teaches the transgene sequence encodes a genome-targeting protein as in instant claim 56.
It would have been obvious to one of ordinary skill in the art before the effective filing date to have modified the clade E variant capsid AAV8 or AAVrh.10 protein, nucleic acid encoding the variant capsid protein, and vectors thereof used for retrograde transport of North Carolina by inserting the AAV variant insert sequence comprising LADQDYTKTA of Hwang into the clade E variant capsid AAV8 or AAVrh.10 protein of North Carolina, to enhance retrograde transport with a reasonable expectation of success. There would be a reasonable expectation of success because North Carolina teaches both AAV8 and AAVrh.10 capsid proteins (Clade E) with modifications and that AAVrh.10 is known to efficiently cross the BBB and taught virus vectors used for retrograde transport, and because Hwang taught while the capsid polypeptide of a virus particle can contain a consensus sequence as described herein, the remaining components can be from one or more known AAV serotypes (e.g. AAV2, AAV8, etc.). Therefore, one of ordinary skill in the art would have been motivated to do so because the AAV8 and AAVrh.10 Clade E AAV capsid proteins, nucleic acids, and vectors thereof of North Carolina were ready for further modification and can be used for retrograde transport and Hwang taught consensus sequences were identified that when present in an AAV capsid protein result in a profound enhancement in the efficacy of retrograde transport, including the sequence LADQDYTKTA as an exemplary insertion into AAV8 to enhance retrograde movement.
Accordingly, the limitations of claims 1,5 and 7 would have been prima facie obvious to one of ordinary skill in the art before the effective filing date.
It would have been obvious to one of ordinary skill in the art to provide a recombinant AAV particle comprising the modified AAV clade E variant capsid AAV8 or AAVrh.10 protein of North Carolina modified by the insert sequence of Hwang et al. with a nucleic acid encoding a transgene sequence based on the teachings of North Carolina and Hwang with a reasonable expectation of success. There would be a reasonable expectation of success because North Carolina taught a virus (e.g., AAV) particle that functions as a nucleic acid delivery vehicle, and which comprises the vector genome packaged within a virion and taught an AAV1/rh.10 chimera capable of crossing the blood-brain barrier containing a CBh-scGFP transgene, and Hwang taught viral particles include a viral capsid protein as described herein and that the viral particle exhibits a preference for retrograde movement and the viral particle possesses retrograde transport capability, and further includes a nucleic acid encoding a payload. One of ordinary skill in the art would have been motivated to do so because Hwang taught that the viral particle exhibits a preference for retrograde movement and the viral particle possesses retrograde transport capability, and that the payload could be a protein-coding gene and an inhibitory RNA nucleic acid, an effector protein including a gene editing system (e.g., CRISPR/Cas9, TALEN, zinc finger nuclease) and exemplifies SaCas9 in AAV2-retro.
Accordingly, the limitations of claims 15,16 and 56 would have been prima facie obvious to one of ordinary skill in the art before the effective filing date.
Claims 2 and 3 are rejected under 35 U.S.C. 103 as being unpatentable over North Carolina in view of Hwang as applied to claims 1,5,7,15,16 and 56 above, and further in view of He et al. (WO 2022098893, EFD 06 Nov 2020).
The teachings of North Carolina and Hwang as applicable to claims 1,5,7,15,16 and 56 has been described above.
North Carolina and Hwang do not teach that the LADQDYTKTA sequence immediately follows a QQQN sequence, or an amino acid sequence having 85% or more sequence identity with the amino acid sequence set forth in SEQ ID NO: 33.
Before the effective filing date, He et al. taught viral vectors and methods for using them to treat injuries and diseases affecting the central nervous system, and a method for targeted retrograde infection of an injured neuron (Page 1, lines 20-24). He et al. taught a VP1 which is AAV10Retro VP1 containing the amino acid sequence LADQDYTKTA (page 4, lines 5-7).
He et al. taught an AAV10 Retro VP1 polypeptide sequence (page 10) which has 98.4% identity to instant SEQ ID NO: 33, and includes the LADQDYTKTA sequence at positions 551-600 which follows a “QQQN” sequence and the sequence alignment is shown below:
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He et al. taught other AAV serotypes useful in the vectors and methods of this disclosure include AAV8, AAV10, rh.10, and AAV7, AAV9, and AAV10 capsids can facilitate targeted crossing of the blood-brain barrier (BBB) or the blood spinal cord barrier (BSCB) at a site of nerve cell injury or damage following intravenous administration to transduce a neuron (page 42, lines 21-28).
It would have been obvious to one of ordinary skill in the art before the effective filing date to have substituted the clade E variant capsid AAV8 or AAVrh.10 protein used for retrograde transport of North Carolina modified by inserting the AAV variant insert sequence comprising LADQDYTKTA of Hwang with the AAV10 Retro VP1 sequence of He et al. to use for targeted retrograde infection of an injured neuron. There would be a reasonable expectation of success this amounts to simple substitution of one known element for another to obtain predictable results. One of ordinary skill in the art would have been motivated to do so because He et al. taught AAV capsid proteins for targeted retrograde infection of an injured neuron and using them to treat injuries and diseases affecting the central nervous system and that other AAV serotypes useful in the vectors and methods of this disclosure include AAV8, AAV10, rh.10, and AAV7, AAV9, and AAV10 capsids can facilitate targeted crossing of the blood-brain barrier (BBB) or the blood spinal cord barrier (BSCB) at a site of nerve cell injury or damage following intravenous administration to transduce a neuron. He et al. taught an AAV10 Retro VP1 polypeptide sequence (page 10) which has 98.4% identity to instant SEQ ID NO: 33, and includes the LADQDYTKTA sequence at positions 551-600 which follows a “QQQN” sequence and therefore which meets the structural limitations of instant claims 2 and 3.
Accordingly, the limitations of claims 2 and 3 would have been prima facie obvious to one of ordinary skill in the art before the effective filing date.
Claim 17 is rejected under 35 U.S.C. 103 as being unpatentable over North Carolina in view of Hwang as applied to claims 1,5,7,15,16 and 56 above, and further in view of Liang et al. (Nature Communications, 7:13282, Published 31 Oct 2016).
The teachings of North Carolina and Hwang as applicable to claims 1,5,7,15,16 and 56 has been described above. Hwang taught viral particles including a nucleic acid encoding a payload (transgene) which may be a protein-coding gene or an inhibitory RNA nucleic acid (page 3, lines 18-28).
North Carolina and Hwang do not teach the transgene sequence encodes a short hairpin RNA that targets CamKv.
Liang et al. taught Ca2þ/calmodulin-dependent protein kinases (CaMKs) are an important class of Ca2þ effectors that trigger synaptic structural modifications (page 1, left column) and that CaMKv synthesis is induced by neuronal activity and sensory experience, and CaMKv is required for activity-dependent dendritic spine maintenance. In addition, CaMKv regulates spine morphogenesis in a Ca2þ/calmodulin-dependent manner (page 2, right column). Liang et al. taught the subcellular localization of CaMKv mRNA and protein as well as the tight regulation of CaMKv expression by synaptic activity suggest that it may play a key role at synapses. As CaMKs are implicated in spine morphogenesis, Liang et al. determined whether the depletion of CaMKv affects dendritic spine density and morphology. Two short hairpin RNAs (shRNAs) designed for CaMKv knockdown in cultured neurons were effective, and CaMKv knockdown by either shRNA significantly reduced dendritic spine density (page 4, left column).
Liang et al. taught changes in spine density are closely correlated with synaptic transmission efficacy. To determine whether CaMKv knockdown regulates functional connectivity between neurons, the frequency and amplitude of miniature excitatory postsynaptic currents (mEPSCs) were measured in dissociated hippocampal neurons on CaMKv knockdown. Consistent with spine density reduction, CaMKv knockdown reduced mEPSC frequency without affecting amplitude, and therefore Liang et al. found that CaMKv is essential for spontaneous excitatory neurotransmission (pages 4-5).
It would have been obvious to one of ordinary skill in the art before the effective filing date, to provide an shRNA targeting CamKv as the transgene in the recombinant AAV particle of North Carolina in view of Hwang in order to determine the effect of knocking down CamKv using shRNA based on the teachings of Liang et al. There would be a reasonable expectation of success, because Hwang et al. taught the transgene could be an inhibitory nucleic acid and therefore would amount to simple substitution of an inhibitory nucleic acid for an shRNA targeting CamKv. One of ordinary skill in the art would have been motivated to do so in order to determine the function and role of CaMKv as Liang et al. taught determining whether the depletion of CaMKv affects dendritic spine density and morphology using shRNA targeting CaMKv, and that CaMKv knockdown by either shRNA significantly reduced dendritic spine density and also reduced mEPSC frequency without affecting amplitude, and therefore Liang et al. found that CaMKv is essential for spontaneous excitatory neurotransmission.
Accordingly, the limitations of claim 17 would have been prima facie obvious to one of ordinary skill in the art before the effective filing date.
Claims 8,54 and 55 are rejected under 35 U.S.C. 103 as being unpatentable over North Carolina in view of Hwang as applied to claims 1,5,7,15,16 and 56 above, and further in view of Gao et al. (US 20030228282, Published 11 Dec 2003).
The teachings of North Carolina and Hwang as applicable to claims 1,5,7,15,16 and 56 has been described above. Hwang taught viral particles including a nucleic acid encoding a payload (transgene) which may be a protein-coding gene or an inhibitory RNA nucleic acid (page 3, lines 18-28).
North Carolina and Hwang do not teach that the transgene sequence is flanked by inverted terminal repeat sequences (ITRs).
Before the effective filing date, Gao et al. taught novel AAV sequences, compositions containing these sequences, and uses therefore including that these compositions are particularly well suited for use in compositions requiring re-administration of rAAV for therapeutic or prophylactic purposes (paragraph 0005). Gao et al. taught AAV8 sequences and fragments thereof are useful in production of rAAV, and are also useful as antisense delivery vectors, gene therapy vectors, or vaccine vectors and the invention further provides nucleic acid molecules, gene delivery vectors, and host cells which contain the AAV8 sequences of the invention (paragraph 0011). Gao et al. taught vectors containing sequences encoding an AAV8 capsid or fragment thereof, and that vectors of the invention further contain a minigene comprising a selected transgene which is flanked by AAV 5’ ITR and AAV 3’ ITR (paragraph 0041) and the minigene is composed of, at a minimum, a transgene and its regulatory sequences, and 5′ and 3′ AAV inverted terminal repeats (ITRs) (paragraph 0050).
It would have been obvious to one of ordinary skill in the art before the effective filing date to further modify the nucleic acid or vector encoding the clade E variant capsid AAV8 or AAVrh.10 protein of North Carolina modified by the insert sequence comprising LADQDYTKTA of Hwang and which comprises a transgene which is a protein-coding gene or an inhibitory RNA nucleic acid, based on the teachings of Gao et al. to comprise ITR sequences flanking the transgene. There would be a reasonable expectation of success, because both North Carolina and Hwang et al. teach AAVs containing transgenes and Gao also taught AAV8 sequences and vectors containing transgenes. One of ordinary skill in the art would have been motivated to do so because Gao et al. taught AAV8 sequences useful in production of rAAV, useful as antisense delivery vectors, gene therapy vectors, or vaccine vectors and the invention further provides nucleic acid molecules, gene delivery vectors which contain the AAV8 sequences of the invention, and taught vectors containing sequences encoding an AAV8 capsid or fragment thereof, and that vectors of the invention further contain a minigene comprising a selected transgene which is flanked by AAV 5’ ITR and AAV 3’ ITR (paragraph 0041) and the minigene is composed of, at a minimum, a transgene and its regulatory sequences, and 5′ and 3′ AAV inverted terminal repeats (ITRs) (paragraph 0050).
Accordingly, the limitations of claims 8,54 and 55 would have been prima facie obvious to one of ordinary skill in the art before the effective filing date.
Claim 10 is rejected under 35 U.S.C. 103 as being unpatentable over North Carolina in view of Hwang and further in view of Gao et al. as applied to claims 8,54 and 55 above, and further in view of Liang et al. (Nature Communications, 7:13282, Published 31 Oct 2016).
The teachings of North Carolina, Hwang and Gao et al. as applicable to claims 8,54 and 55 have been described above. Hwang taught viral particles including a nucleic acid encoding a payload (transgene) which may be a protein-coding gene or an inhibitory RNA nucleic acid (page 3, lines 18-28).
North Carolina and Hwang do not teach the transgene sequence encodes a short hairpin RNA that targets CamKv.
Liang et al. taught Ca2þ/calmodulin-dependent protein kinases (CaMKs) are an important class of Ca2þ effectors that trigger synaptic structural modifications (page 1, left column) and that CaMKv synthesis is induced by neuronal activity and sensory experience, and CaMKv is required for activity-dependent dendritic spine maintenance. In addition, CaMKv regulates spine morphogenesis in a Ca2þ/calmodulin-dependent manner (page 2, right column). Liang et al. taught the subcellular localization of CaMKv mRNA and protein as well as the tight regulation of CaMKv expression by synaptic activity suggest that it may play a key role at synapses. As CaMKs are implicated in spine morphogenesis, Liang et al. determined whether the depletion of CaMKv affects dendritic spine density and morphology. Two short hairpin RNAs (shRNAs) designed for CaMKv knockdown in cultured neurons were effective, and CaMKv knockdown by either shRNA significantly reduced dendritic spine density (page 4, left column).
Liang et al. taught changes in spine density are closely correlated with synaptic transmission efficacy. To determine whether CaMKv knockdown regulates functional connectivity between neurons, the frequency and amplitude of miniature excitatory postsynaptic currents (mEPSCs) were measured in dissociated hippocampal neurons on CaMKv knockdown. Consistent with spine density reduction, CaMKv knockdown reduced mEPSC frequency without affecting amplitude, and therefore Liang et al. found that CaMKv is essential for spontaneous excitatory neurotransmission (pages 4-5).
It would have been obvious to one of ordinary skill in the art before the effective filing date, to provide an shRNA targeting CamKv as the transgene in the recombinant AAV of North Carolina in view of Hwang and further in view of Gao et al. in order to determine the effect of knocking down CamKv using shRNA based on the teachings of Liang et al. There would be a reasonable expectation of success, because Hwang et al. taught the transgene could be an inhibitory nucleic acid and therefore would amount to simple substitution of an inhibitory nucleic acid for an shRNA targeting CamKv. One of ordinary skill in the art would have been motivated to do so in order to determine the function and role of CaMKv as Liang et al. taught determining whether the depletion of CaMKv affects dendritic spine density and morphology using shRNA targeting CaMKv, and that CaMKv knockdown by either shRNA significantly reduced dendritic spine density and also reduced mEPSC frequency without affecting amplitude, and therefore Liang et al. found that CaMKv is essential for spontaneous excitatory neurotransmission.
Accordingly, the limitations of claim 10 would have been prima facie obvious to one of ordinary skill in the art before the effective filing date.
Conclusion
Claims 1-3,5,7,8,10,15-17 and 53-56 are rejected.
Claim 53 which requires the entire amino acid sequence of SEQ ID NO: 32 is free of the prior art.
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/STEPHANIE L SULLIVAN/Examiner, Art Unit 1635
/ABIGAIL VANHORN/Primary Examiner, Art Unit 1636
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