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 .
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
Priority
Th instant application, filed 03/10/2023, claims domestic benefit to US provisional applications 63/320,907, filed 03/17/2022, and 63/320,567, filed 03/16/2022.
Status of Claims/Application
Applicant’s preliminary amendment of 05/29/2023 is acknowledged. Claims 3, 5-11, 14, 16, 19, 22-24, 27, 29, 32, 35-37, 41, and 44 are amended and claims 15, 17, 20-21, 25-26, 28, 30, 33-34, 38-40, 42-43, and 45 are cancelled. Claims 1-14, 16, 18-19, 22-24, 27, 29, 31-32, 35-37, 41, and 44 are currently pending.
Election/Restrictions
Applicant’s election without traverse of the following in the reply filed on 10/07/2025 is acknowledged.
Applicant was required to elect a single invention from Groups I (engineered cell claims) or II (method claims). In the response, applicant elected Group I, which reads on instant claims 1-10.
From group I, applicant was further required to elect from the following species:
A single specific transmembrane and juxtamembrane domain. Applicant elected NOTCH1 for both.
A single and specific heterologous recognition motif. Applicant elected collagen type II.
A single and specific therapeutic transgene. Applicant elected ILRa.
Upon further consideration, the restriction requirement between Groups I and II is withdrawn. The species election is maintained.
Claims 1-14, 16, 18-19, 22-24, 27, 29, 31-32, 35-37, 41, and 44 are examined on the merits herein in so far as they apply to the elected species.
Information Disclosure Statement
The information disclosure statement (IDS) submitted on 06/18/2024 is in compliance with the provisions of 37 CFR 1.97, except where noted below. Accordingly, the information disclosure statement has been considered by the examiner.
A copy of NPL reference Ulrich-Vinther, et al (C40) was not found in the file wrapper in accordance with 37 CFR 1.97. The reference has been lined through on the IDS and has not been considered.
Nucleotide and/or Amino Acid Sequence Disclosures
The instant application recites sequences that require a sequence listing, but a sequence listing has not been filed. For instance, see at least the following sequences that comprise four or more amino acids and require an associated SEQ ID NO:
Claim 4: “WYRGRL”;
The instant specification:
page 4, line 23, “WYRGRL”; and
page 7, line 12, “NITEGE”
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 - This application fails to comply with the requirements of 37 CFR 1.821 - 1.825 because it does not contain a "Sequence Listing" as a separate part of the disclosure or a CRF of the “Sequence Listing.”.
Required response - Applicant must provide:
A "Sequence Listing" part of the disclosure; together with
An amendment specifically directing its entry into the application in accordance with 37 CFR 1.825(a)(2);
A statement that the "Sequence Listing" includes no new matter as required by 37 CFR 1.821(a)(4); and
A statement that indicates support for the amendment in the application, as filed, as required by 37 CFR 1.825(a)(3).
If the "Sequence Listing" part of the disclosure is submitted according to item 1) a) or b) above, Applicant must also provide:
A substitute specification in compliance with 37 CFR 1.52, 1.121(b)(3) and 1.125 inserting the required incorporation-by-reference paragraph, 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.
If the "Sequence Listing" part of the disclosure is submitted according to item 1) c) or d) above, applicant must also provide:
A CRF in accordance with 37 CFR 1.821(e)(1) or 1.821(e)(2) as required by 1.825(a)(5); and
A statement according to item 2) a) or b) above.
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).
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.
Drawing Objection
The drawings filed 03/10/2023 and 05/29/2023 are objected to for containing colored drawings without an approved petition for colored drawings. The filing, and approval, of a petition for colored drawings, or replacement drawings in black and white/grayscale, are required.
Color photographs and color drawings are not accepted in utility applications unless a petition filed under 37 CFR 1.84(a)(2) is granted. Any such petition must be accompanied by the appropriate fee set forth in 37 CFR 1.17(h), one set of color drawings or color photographs, as appropriate, if submitted via the USPTO patent electronic filing system or three sets of color drawings or color photographs, as appropriate, if not submitted via the via USPTO patent electronic filing system, and, unless already present, an amendment to include the following language as the first paragraph of the brief description of the drawings section of the specification:
The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.
Color photographs will be accepted if the conditions for accepting color drawings and black and white photographs have been satisfied. See 37 CFR 1.84(b)(2).
The drawings are further objected to for having low resolution in the following figures making them difficult to read. For instance, see:
Fig 1, specifically the equation that is provided on the top right of the bottom two images;
Fig 2A-B, specifically label text in/for plots; and
Fig. 4
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.
Claim Objections
Claim 2 is objected to for the following informalities: the claim recites the following juxtamembrane domains twice: NOTCH1 (lines 4 and 5) and PTPRF (line 5). Correction to remove the redundant recitation is suggested.
Claim 3 is objected to for the following informality: line 4 of the claim recites “Ga14VP64”. The “1” (one) that is following the “Ga” appears to be a typo for the letter “l” and the recitation should be “Gal4VP64”. For instance, see the recitation of “Gal4VP64, in the instant specification, page 4, line 18.
Claim Rejections - 35 USC § 112(b)
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 2-6 and 16 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.
Claim 2, line 2 recites transmembrane domains including “Notch4” and “Notch1” while lines 4-5 of the claim recites juxtamembrane domains including “NOTCH1” and “NOTCH4”. It is unclear if the differences in capitalization, for example “Notch1” versus “NOTCH1” are intended to reference different structures, for instance a gene versus a protein or the domain of a specific species, or if the recited domains reference the same thing and are just inconsistently written.
Appropriate correction/clarification is required.
In the instant office action, the nomenclature used in the claim is not interpreted to be structurally limiting, for instance of a gene vs a protein, and the claim is interpreted as encompassing any NOTCH4 or NOTCH1 transmembrane domain from any species in either gene or protein form.
Regarding claims 2-4, the phrase "such as" in the following recitations renders the claims indefinite because it is unclear whether the limitations following the phrase are part of the claimed invention. See MPEP § 2173.05(d).
Claim 2, line 2 and line 4, “such as one selected from…”;
Claim 3, line 2, “such as where a Notch extracellular domain is replaced…”
Claim 3, line 4, “such as Ga14VP64 or the tetracycline transactivator”
Claim 4, line 2, “such as an scFv, a nanobody, or…”
Appropriate correction is required.
Claim 4 recites the limitation "the heterologous recognition motif" in lines 1 and 3. There is insufficient antecedent basis for this limitation in the claim. Claim 4 depends on claim 2 and ultimately claim 1. The claims do not recite a heterologous recognition motif that could be being referenced.
Appropriate correction is required.
In the instant office action, the claim is interpreted as being dependent on claim 3, which recites a heterologous recognition motif.
Regarding claim 6, lines 3-5 of the claim recites “CRISPR-based transcriptome modulators (e..g. such as Rfx Cas13D, dCas9-VPR, dCas9-KRAB or CRISPRoff/DNMT3A-DNTM3L-dCas9-KRAB fusion protein or CRISPRon/TET1-XTEN80-dCas9 fusion protein).” The use of “e.g.” and “such as” as well as the inclusion of the limitations within parentheticals renders the claim indefinite as it is unclear whether the limitations following the phrases/included in parenthesis are part of the claimed invention or exemplary embodiments. See MPEP 2173.05. This is particularly the case as the limitations in parenthesis are narrower embodiments of the preceding limitation.
A broad range or limitation together with a narrow range or limitation that falls within the broad range or limitation (in the same claim) may be considered indefinite if the resulting claim does not clearly set forth the metes and bounds of the patent protection desired. See MPEP § 2173.05(c). In the present instance, claim 6 recites the broad recitation CRISPR-based transcriptome modulators, and the claim also recites (e..g. such as Rfx Cas13D, dCas9-VPR, dCas9-KRAB or CRISPRoff/DNMT3A-DNTM3L-dCas9-KRAB fusion protein or CRISPRon/TET1-XTEN80-dCas9 fusion protein) which is the narrower statement of the range/limitation. The claim(s) are considered indefinite because there is a question or doubt as to whether the feature introduced by such narrower language is (a) merely exemplary of the remainder of the claim, and therefore not required, or (b) a required feature of the claims.
Claim 3 contains the trademark/trade name “synNotch”. Where a trademark or trade name is used in a claim as a limitation to identify or describe a particular material or product, the claim does not comply with the requirements of 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph. See Ex parte Simpson, 218 USPQ 1020 (Bd. App. 1982). The claim scope is uncertain since the trademark or trade name cannot be used properly to identify any particular material or product. A trademark or trade name is used to identify a source of goods, and not the goods themselves. Thus, a trademark or trade name does not identify or describe the goods associated with the trademark or trade name. In the present case, the trademark/trade name is used to identify/describe a synthetic signaling receptor derived from the Notch receptor and, accordingly, the identification/description is indefinite.
Claim 16 recites “The method of claim 1”. There is insufficient antecedent basis for this limitation in the claim. Claim 1 is drawn to a product (an engineered cell), not a method and does not recite a method that could be being referenced rendering the metes and bounds of claim 16 indefinite.
Appropriate correction is required.
In the instant office action, the claim is interpreted as being dependent on claim 11, which is drawn to a method.
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.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
Claims 1, 3-4, 6, and 8 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Cho, J.H., et al (2018) Engineering Axl specific CAR and SynNotch receptor for cancer therapy Scientific Reports 8(3846); 1-8.
Cho teaches that the transfer of engineered T cells expressing chimeric antigen receptors (CAR) is an exciting cancer therapeutic approach that shows high efficacy against cancers in clinical trials, especially for B cell malignancies. Furthermore, recently developed synthetic NOTCH (synNOTCH) receptor has demonstrated potential in enhancing the specificity of CAR T cell therapy and delivering therapeutic payloads to tumors in an antigen-dependent manner. Cho developed a single-chain variable fragment (scFv) from a humanized monoclonal antibody against Axl and used the scFv in an anti-Axl synNotch domain (abstract).
Cho teaches that a novel receptor design called synNotch had been created that enables both the programming of input and output via the release of intracellular transcription factor upon antigen-receptor binding. The synNotch receptor uses the regulatory notch core portion with an engineered transcription factor that enables programmable inputs and outputs to perform user-defined functions. Because of the high programmability, synNotch has been used to reprogram human primary T cell responses both in vitro and in vivo for enhancing tumor specificity and delivering therapeutic payloads in a tumor antigen-specific manner. As such, synNotch receptor targeting Axl ligand with different output functions, such as producing a defined set of cytokines, will improve cellular immunotherapy to treat various cancers. In this study, Cho engineered an Axl synNotch receptor for producing IL-10 in an antigen-specific manner (page 2, paragraph 1).
Fig. 4 of Cho (page 5) shows the design and characterization of the Axl synNotch in human Jurkat T cells. Fig 4a is duplicated below for convenience, where TF is a transcription factor and TFBD is a transcriptional factor binding domain (Fig. 4 description, page 5):
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Cho teaches that the Axl synNotch receptor is composed of the Axl scFv as the extracellular domain and notch core region fused to the engineered transcription factor tTA. Jurkat T cells were engineered to stably express the synNotch receptor using electroporation and piggyBac transposon-based system. A reporter construct that composes of a tTA responsive promoter followed by a gene of interest (e.g., blue fluorescent protein (BFP) or IL-10) were also introduced into the Jurkat T cells. IL-10 was chosen as an output because IL-10 is an inhibitory cytokine that can be used to control inflammation. When Axl synNotch expressing T cells engage with tumor cells that express Axl, the tTA transcription factor is cleaved from the synNotch and translocated into the nucleus to regulate gene expression from the reporter (paragraph bridging pages 3-4).
Cho demonstrates the therapeutic potential of Axl scFv by designing a functional Axl synNotch receptor that can produce IL-10 when activated by Axl+ tumor cells (page 6, conclusion).
Thus, Cho anticipates instant claims 1, 3-4, 6, and 8.
Claims 1-4, 6, 8, and 10 are rejected under 35 U.S.C. 102(a)(1) and (a)(2) as being anticipated by US 10,836,808 B2 (Lim, W.A., et al) 17 NOV 2020.
US’808 teaches a molecular circuit comprising:
(a) a nucleic acid encoding a chimeric notch receptor polypeptide comprising:
(i) an extracellular binding domain that comprises an antigen binding region of an antibody;
(ii) a notch receptor regulatory domain; and
(iii) an intracellular domain that comprises a DNA binding domain,
wherein, binding of the antigen binding region of (a)(i) to an antigen induces proteolytic cleavage of the Notch receptor regulatory domain of (a)(ii) and releases the DNA binding domain of (a)(III); and
(b) an expression cassette comprising a transcriptional control element operably linked to a nucleic acid encoding a therapeutic agent,
wherein, the released DNA binding domain binds to the transcriptional control element and induces expression of the nucleic acid encoding the therapeutic agent (col. 187, claim 1).
US’808 also teaches host cells genetically modified with the nucleic acids (abstract).
US’808 teaches binding triggered transcriptional switch polypeptides, nucleic acid sequences encoding the polypeptides, and host cells genetically modified with the nucleic acids (Col. 1, lines 55-60). US’808 teaches that notch receptors are transmembrane proteins that mediate cell-cell contact signaling and play a central role in development and other aspects of cell-cell communication. Notch receptors expressed in a receiver cell recognize their ligands (the delta family of proteins), expressed on a sending cell. The engagement of notch and delta on the contacting cells leads to two-step proteolysis of the notch receptor that ultimately causes the release of the intracellular portion of the receptor from the membrane into the cytoplasm. This released domain alters receiver cell behavior by functioning as a transcriptional regulator (Col. 1, lines 35-53).
US’808 teaches synthetic notch receptors as a modular platform for engineering customized cell sensing and response behaviors. A minimal notch transmembrane region can be combined with novel extracellular and intracellular domains to construct diverse chimeric receptors. The core of the notch transduction pathway is the transmembrane region. This design can be used as a platform for engineering a series of receptors that detect a diverse array of membrane-resented ligands. On the intracellular side, the notch intracellular domain (NICD) can be replaced by orthogonal transcription factors to report on notch signaling activation. On the extracellular, the endogenous delta binding domain can be replaced by a protein binding domain, for example FKBP, an anti-GFP nanobody, etc. and cleave is induced when the cognate binding partner is bound. Therefore, libraries of receptor molecules with different extracellular domains coupled with a different intracellular domain were generated as shown in Figs. 37 and 38 (col. 152, lines 35-60). Fig. 37B is duplicated below for convenience:
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As shown, the synNotch modules generated by US’808 include scFvs or nanobodies as well as intracellular domains including Gal4-VP64, or tTa. The synNotch core comprises the 3 Lin12-Notch repeats, the heterodimerization domain, and the transmembrane of Notch, which is the juxtamembrane domain in combination with the transmembrane domain of Notch.
The teachings of US’808 also indicate that the Notch domains are from Notch 1, as shown in Fig. 2A (col. 38, lines 40-67).
US’808 exemplifies engineered T cells with customized therapeutic response programs using synthetic notch receptors. In the study, synNotch receptors were built by fusing the CD19 scFv, LaG17 (lower affinity), or LaG16_2 (high affinity GFP nanobody to the mouse Notch1 (NM_008714) minimal regulatory region and Gal4VP64. All synNotch receptors contained a n-terminal CD8a signal peptide for membrane targeting and a myc-tag for easy determination of surface expression. Human IL-2, IL-10, Tbet, or TRAIL codon optimized mRNA sequences were cloned into a MCS downstream of the Gal4 inducible promoter and 5’ of an IRES mCherry reporter (col. 160, line 44 – col. 161, line 4). Primary CD4+ and CD8+ T cells were isolated from anonymous donor blood and were transduced using lentiviral transduction (col. 161, lines 5-34). Cytokine production was then evaluated using cancer cell lines as sending cells for in vitro stimulation of synNotch T cells (col. 161, line 35 – col. 162, line 67). US’808 also describes animal studies with an anti-CD19 targeting synNotch receptor using Gal4VP64 and response elements controlling human IL-2 expression (col. 163, line 1 – col. 164, line 53). As US’808 teaches studies in mice, the cells would necessarily have been in a pharmaceutical formulation.
Thus, US’808 anticipates instant claims 1-4, 6, 8, and 10.
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 1-6, 8-11, 13, 16, 18-19, 23, 29, 32, and 36-37 are rejected under 35 U.S.C. 103 as being unpatentable over Glass, K.A., et al (2014) Tissue-engineered cartilage with inducible and tunable immunomodulatory properties Biomaterials 35; 5921-5931 in view of Morsut, L., et al (2016) Engineering customized cell sensing and response behaviors using synthetic Notch receptors Cell 164; 780-791 and Supplementary materials and Sulaiman, S.B., et al (2021) Type II collagen-conjugated mesenchymal stem cells micromass for articular tissue targeting Biomedicines 9(880); 1-17 as evidenced by Sulaiman, S.B., et al (2020) Gelatin microsphere for cartilage tissue engineering: current and future strategies Polymers 12(2404); 1-16 (herein “Sulaiman2020”).
Glass teaches that pathogenesis of osteoarthritis is mediated in part by inflammatory cytokines including interleukin-1 (IL-1), which promote degradation of articular cartilage and prevent human mesenchymal stem cell (MSC) chondrogenesis. Glass combines gene therapy and functional tissue engineering to develop engineered cartilage with immunomodulatory properties that allow chondrogenesis in the presence of pathologic levels of IL-1 by inducing overexpression of IL-1 receptor antagonist (IL-1Ra) in MSCs via scaffold mediated gene delivery. A doxycycline inducible vector was used to transduce MSCs in monolayer or within 3D woven PCL scaffolds to enable tunable IL-1Ra production. In the presence of IL-1, IL-1Ra expressing engineered cartilage produced cartilage-specific extracellular matrix, while resisting IL-1 induced upregulation of matrix metalloproteinases and maintaining mechanical properties similar to native articular cartilage. The ability of functional engineered cartilage to deliver tunable anti-inflammatory cytokines to the joint may enhance the long-term success of therapies for cartilage injuries or osteoarthritis (abstract).
Glass teaches that the pathogenesis of OA and post-traumatic arthritis following joint injury, is mediated in part by the action of pro-inflammatory cytokines such as IL-1, which are found at elevated concentration in the synovial fluid of OA joints. More recent evidence shows that IL-1 also prevents MSC chondrogenesis and matrix accumulation in pellet culture and within biomaterial scaffolds. In this regard, in vivo implantation of MSCs for articular cartilage repair can be associated with a loss of chondrogenic potential as well as a shift toward a more hypertrophic phenotype, which may result in endochondral ossification of the implant. There is growing evidence that the inflammatory environment within the joint may be in part responsible for this altered MSC differentiation. Thus, inflammatory signaling mediated by IL-1 within the OA or the injured joint may inhibit the development and homeostasis of tissue engineered cartilage while continuing to degrade native tissue (paragraph bridging pages 5921-5922).
IL-1Ra is a natural inhibitor of IL-1 that competes with IL-1 in binding to the IL-1 receptor. Daily systemic injection of recombinant human IL-1Ra (anakinra) is approved for treatment of rheumatoid arthritis and has been explored in OA treatment, but its efficacy is limited by its short half-life of only a few hours. Intra-articular gene delivery of IL-1Ra, or delivery of cells that have been transduced ex vivo, has been studied extensively in animal models and has progressed to clinical trials. To date, IL-Ra gene therapy strategies for OA have employed constitutive expression cassettes, which lack regulation of transgene expression (page 5922, left column, paragraph 2).
Glass teaches MSCs seeded into scaffolds were efficiently engineered to express transgenes, leading to robust chondrogenic differentiation without the need to supplement cultures with exogenous growth factors (page 5922, left column, paragraph 3). Glass employed scaffold-mediated LV gene delivery of a dox-inducible IL-1Ra expression vector to MSCs to engineer cartilage constructs with tunable IL-1Ra overexpression (page 5922, paragraph bridging columns).
Glass demonstrates that the engineered cartilage constructs with differentiating human MSCs expressed PGE2 during chondrogenesis in the absence of IL-1. This finding is consistent with previous studies showing human MSCs expressing PGE2 during chondrogenesis in pellet culture. In the current study, PGE2 was undetectable at days 18 and 27 in IL-1 treated control constructs, possibly due to the lack of chondrogenic differentiation in these samples. IL-1Ra expressing constructs, in contrast, maintained similar PGE2 levels to those of untreated constructs. PGE2 is also important for chondrogenic commitment of progenitors during endochondral ossification and contributes to the immunomodulatory properties of undifferentiated MSCs in vivo (page 5928, right column, paragraph 4).
Glass further teaches enhanced collagen production in IL-1Ra expressing constructs under inflammatory conditions. In the absence of IL-1, engineered cartilage constructs harvested at day 27 accumulated total collagen normalized to DNA (collagen/DNA) of ~50 μg/μg. Collagen was not detectable at the time of seeding or chondrogenic induction. Treatment with IL-1 significantly reduced collagen deposition in NT and eGFP expressing constructs by ~2 fold, with similar levels observed at both doses of IL-1. IL-1Ra expressing constructs were protected from IL-1 mediated reduction of collagen content, showing similar total collagen deposition to that of constructs in the absence of IL-1. Similar trends were observed for total collagen content per construct without normalizing to DNA. Furthermore, immunohistochemistry (IHC) was performed to examine the deposition of cartilage-specific types of collagen. Type II collagen is the most abundant collagen found in articular cartilage and IHC revealed strong type II collagen staining in untreated constructs, which was maintained in IL-1Ra expressing constructs treated with IL-1 (paragraph bridging pages 5925-5926).
While Glass teaches methods of increasing MSC chondrogenic differentiation, which reasonably be expected to directly result in an increase in cartilage mass/volume, by engineering MSC to over express IL-1Ra, Glass does not disclose that the MSCs comprise a synthetic receptor that functions as recited in claim 1.
Morsut teaches that customized cell sensing/response pathways would be extremely useful for engineering therapeutic cells, allowing them to autonomously sense user-specific disease or injury signals, and precisely deploying therapeutic or repair functions (page 780, left column, paragraph 1). Morsut teaches that, to generate synthetic pathways that allow for customizable sensing and response engineering, the Notch pathway was used, because of its very direct and simple mechanism of transduction. Engagement of the Notch receptor with its ligand leads to intermembrane proteolysis. The induced cleave of the receptor releases the intracellular fragment of Notch. This Notch intracellular domain is a transcriptional regulator that can only function when it is released from the membrane and can enter the nucleus to activate target genes that play key roles in cell-cell signaling during development (page 780, right column, paragraph 3).
Morsut teaches that, in native Notch, the LNR domains mask the protease cleavage site in the unbound conformation. When the ligand engages the receptor, mechanical force is thought to expose this protease site, initiating the multi-step process leading to release of the intracellular transcription domain. Morsut proposes that when the extracellular domain is replaced by a novel recognition domain, the proper cell-cell engagement of the cognate ligand can exert a similar force to expose the Notch core to proteolysis (page 789, Fig. 7, and Fig. 7 caption).
Morsut further teaches that prior studies had shown that the intracellular domain of Notch can be replaced with an artificial transcription factor, for example Gal4-VP64, to create a reporter of Notch activity (page 780, right column, paragraph 4). Studies of the physical mechanism of Notch activation have also shown that the extracellular domain of Notch can be replaced by alternative domains. This very direct mechanism of Notch has inspired the engineering of novel proteolytically induced receptors and reporter systems (page 781, left column, paragraph 2).
Morsut shows that input sensing can be customized by swapping the extracellular recognition domain of these receptors, including the use of antibody based domains, such as single-chain antibodies or nanobodies, to detect a wide range of user specified cell-surface proteins, such as disease antigens. Simultaneously, these novel inputs can be linked to customized responses by swapping the intracellular transcription domain and providing specific downstream effector target genes. The resulting synthetic Notch (synNotch) receptors only retain the minimal transmembrane core domain of the native Notch, which mediates control of proteolysis. These synNotch receptors function in a range of cells (page 781, left column, paragraph 2).
A schematic of the synNotch receptor is provided in Fig. 1 A and B of Morsut. The synNotch receptor schematic from figure 1A is duplicated below for convenience and demonstrates the mechanism of the synNotch receptor in which a target molecule engages the recognition domain resulting in cleavage and the release of the intracellular signaling domain which generates transcriptional output.
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Figure 1B demonstrates that the extracellular domain can be an scFv or a nanobody and the intracellular domain can be Gal4-VP64, TetR-VP64, ZFHD1-VP64, or Gal4-KRAB. Morsut also teaches the use of a tTA intracellular transcription activation domain (page 786, Fig. 5 and Fig. 5 description), which is tetracycline transactivator. Fig. 1B of Morsut is duplicated below:
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As shown in Fig. 1B, the synNotch core comprises the 3 Lin12-Notch repeats (LNR) and a heterodimerization domain of Notch, which is the juxtamembrane domain in combination with the transmembrane domain of Notch. Additionally, Morsut teaches that the cells are provided specific downstream effector target genes, indicating the further inclusion of transgenes. This is also supported by, for instance, Fig. 2, where a GFP reporter gene is expressed upon cleavage of the synNotch core.
Morsut teaches that synNotch receptors were built by fusing the scFvs or nanobodies to the mouse Notch1 (NM_008714) minimal regulatory region (Ile1427 to Arg1752) or extended regulatory region (pro1390 to Arg1752). All receptors contained a N-terminal CD8a signal peptide for membrane targeting an a myc-tag (page 789, left column, paragraph 4).
Morsut also teaches that multiple synNotch pathways can be deployed in the same cell, and can be used to engineer complex combinatorial sensing circuits because they have distinct intracellular and extracellular domains and function orthogonally to one another with no common signaling intermediates (paragraph bridging columns, page 781).
Sulaiman teaches that osteoarthritis (OA) is a chronic degenerative disease of the joints characterized by articular cartilage degeneration of the knee, hip, or hand; synovitis; and the loss of extracellular matrix, accompanied by progressive pain and functional impairment. Treatment of OA depends on its severity, and varies from conservative treatment to invasive surgical intervention such as joint replacement. However, due to the risk of failure and morbidity in the conventional treatments, the potential applicability of cell therapy and tissue engineering for treating OA have been explored (page 1, paragraph 1).
Sulaiman teaches that cell-based therapies for the treatment of OA are not foreign; numerous studies have reported beneficial effects. The therapies feature delivery of cells, particularly mesenchymal stem cells (MSCs) to the knee by means of direct injection or implantation. The desired effects of MSCs therapy in the treatment of OA knees were mostly for its immunomodulatory properties in which it was known that MSC based therapies were approved to be used in graft vs host disease for immunosuppression (paragraph bridging pages 1-2). Sulaiman teaches that previously observed cell apoptosis following intra-articular injection is expected due to several reasons; first, the inhabitable inflammatory environment of an OA knee, and second, the inability of MSCs to survive single-handedly in suspension (page 2, paragraph 3).
Sulaiman studied the effect of palmitated protein G (PPG) intercalation and antibody conjugation on MSCs. Here, protein G was first palmitated (producing PPG), followed by intercalation into the MSC membrane. The second step involved conjugating type II collagen antibody to the PPG-MSCs. Type II collagen is commonly found in the extracellular matrix of cartilage; hence the antibody conjugation would provide high affinity binding of MSCs to the articular defects. The proliferation and differentiation potentials of both PPG-MSCs and antibody-conjugated PPG-MSCs (PPG-MSCs-Ab) in the monolayer and 3D states (GM) were tested. Finally, a preliminary proof of concept of the efficacy of targeted delivery of the PPG-MSC-Ab micromass to an osteochondral defect explant was tested in vivo (paragraph bridging pages 2-3).
Sulaiman teaches that, in monolayer cultures, MSCs were able to proliferate despite being conjugated to PPG. Additionally, in terms of stemness properties, the PPG-MSCs were able to maintain their multi-lineage differentiation to osteocyte, chondrocyte, and adipocyte (page 8, paragraph 2-3).
Sulaiman demonstrates that, after 48 hours of incubation of MSCs onto human osteochondral defect explants, the Ab coated MSCs could be seen attaching on the osteochondral surface. Figure 7 demonstrates a clear distinction in binding capacity between the group injected with MSCs alone and GM-MSC or GM-PPG-MSCs-Ab. There is a significant increase in green fluorescence intensity in GM-PPG-MSCs-Ab compared to GM-MSCs (page 13, paragraph 1; Fig. 7).
Sulaiman further teaches that factors such as high cell density and chondrocyte phenotype maintenance are vital for ensuring successful tissue regeneration upon delivery. Chondrocytes may undergo differentiation and lose their ability to produce hyaline cartilage for tissue restoration; they might instead form the undesirable fibrocartilage. Sulaiman demonstrates that the preservation of the MSCs ability to proliferate and differentiate chondrogenically in both monolayer and 3D culture. In addition, the disclosed findings also confirm the increase of chondrogenic effects by MSCs in the 3D culture as seen in previous studies (page 15, paragraph 2).
Sulaiman concludes that antibody-conjugated MSCs in GM culture is a potential method for targeted delivery of MSCs in future therapy of cartilage defects (abstract).
It would have been prima facie obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to modify the MSC and method of treating osteoarthritis taught by Glass by substituting the Dox-inducible vector disclosed by Glass for the expression of IL-1Ra with a synthetic Notch (synNotch) receptor as disclosed by Morsut where the effector domain of the receptor leads to the expression of IL-1Ra when the receptor core is cleaved. It would have further been obvious to substitute the extracellular binding domain scFvs or nanobodies disclosed by Morsut in the synNotch receptor with a binding domain that targets type II collagen as disclosed by Sulaiman.
An ordinarily skilled artisan would have been able to substitute the dox inducible vector disclosed by Glass with the synthetic Notch receptor disclosed by Morsut with a reasonable expectation of success as the art identifies both as a means for controlled expression of a transgene and both use external cues to control the expression of a transgene. An ordinarily skilled artisan would have been motivated to use an extracellular domain that binds type II collagen as the extracellular domain on the receptor as Sulaiman teaches that type II collagen is commonly found in the extracellular matrix of cartilage, that MSCs with a type II collagen antibody on the surface are able to provide targeted delivery of MSCs to cartilage defects, and teaches that the expression of a type II collagen antibody provides high affinity binding of MSCs to the articular defects in OA. Additionally, these teachings of Sulaiman further motivate the substitution of the dox inducible vector of Glass with the synNotch receptor of Morsut as the additional benefits of direct targeting and high affinity binding to articular defects in OA would be realized. An ordinarily skilled artisan would have had a reasonable expectation of success in using a type II collagen targeting antibody as both Glass and Sulaiman are teaching the treatment of OA using modified MSCs. Additionally, Glass teaches that type II collagen was the most abundant collagen found in the articular cartilage and that IHC revealed strong type II collagen staining in untreated constructs, which was maintained in IL-1Ra expressing constructs treated with IL-1 further demonstrating the presence of type II collagen in the methods taught by Glass.
Regarding claims 23 and 36, the teachings of the combination of Glass, Morsut, and Sulaiman discussed above are with regards to treating osteoarthritis. The combination of applied references do not teach that the patient with OA has cancer and; therefore, the method taught by the combination of applied references meets the claim limitation of the subject not having cancer.
Regarding claim 18, Sulaiman further teaches that a recent study reported that microspheres, specifically gelatin microspheres (GM) increased the chondrogenic differentiation of mesenchymal stem cells (MSCs). As microspheres are of miniscule size, microsphere delivery does not require open surgery, as they easily fit inside a needle for intra-articular injection in humans (page 2, paragraph 4). As discussed above, Sulaiman studied the PPG-MSCs-Ab in the monolayer and in 3D states with GM (page 3, paragraph 1). Sulaiman teaches that the gelatin microspheres were fabricated according to an established method using gelatin dissolved in water then emulsified in olive oil (page 3, paragraph 4). The MSCs were then seeded onto the microspheres (page 4, 2.4). The inter-articular administration of the gelatin microspheres disclosed by Sulaiman meet the instant claim 18 limitation of a time released device that is implanted at the target site as evidenced by Sulaiman2020, which demonstrates that gelatin microspheres prepared with a water in oil emulsification technique provides a controlled release. Sulaiman2020 also discloses that gelatin microspheres are degraded in vitro or in vivo by enzymatic degradation with collagenase, which leads to the controlled release (abstract; page 3, 2.2.2).
It would have been prima facie obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to modify the method taught by the combination of Glass, Morsut, and Sulaiman by administering the engineered MSC using gelatin microspheres administered intra-articular injection as disclosed by Sulaiman. An ordinarily skilled artisan would have been motivated to use the gelatin microspheres of Sulaiman for delivery as Sulaiman demonstrates that microspheres, specifically gelatin microspheres (GM) increased the chondrogenic differentiation of mesenchymal stem cells (MSCs). An ordinarily skilled artisan would have had a reasonable expectation of success as both Glass and Sulaiman teach methods of administering modified MSC for the treatment of osteoarthritis.
Claim 7 is rejected under 35 U.S.C. 103 as being unpatentable over Glass, K.A., et al (2014) Tissue-engineered cartilage with inducible and tunable immunomodulatory properties Biomaterials 35; 5921-5931 in view of Morsut, L., et al (2016) Engineering customized cell sensing and response behaviors using synthetic Notch receptors Cell 164; 780-791 and Supplementary materials and Sulaiman, S.B., et al (2021) Type II collagen-conjugated mesenchymal stem cells micromass for articular tissue targeting Biomedicines 9(880); 1-17 as applied to claim 1 above, and in further view of Wang, Z.H., et al (2014) Delivery of the Sox9 gene promotes chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells in an in vitro model Braz J Med Biol 47(4); 279-286.
The combination of Glass, Morsut, and Sulaiman teach the engineered cell of claim 1 as discussed in detail above.
The combination of applied references, however, does not disclose that the engineered cell comprises a second therapeutic transgene.
Wang teaches that SRY-related high mobility group box 9 (Sox9) gene is a cartilage specific transcription factor that plays essential roles in chondrocyte differentiation and cartilage formation. Wang studied the feasibility of genetic delivery of Sox9 to enhance chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells. After they were isolated from human umbilical cord blood, MSCs were untreated or transfected with a human Sox9 expressing plasmid or empty vector. The cells were assessed for morphology and chondrogenic differentiation. Sox9 overexpression induced accumulation of sulfated proteoglycans without altering cellular morphology. Immunocytochemistry demonstrated that delivery of Sox9 markedly enhanced the expression of aggrecan and type II collagen in MSCs compared with empty vector transfected counterparts. Wang teaches that, taken together, short-term Sox9 overexpression facilitates chondrogenesis of MSCs and may have potential implications in cartilage tissue engineering (abstract).
It would have been prima facie obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to modify the engineered cell disclosed by the combination of Glass, Morsut, and Sulaiman by further including the human Sox9-expressing plasmid disclosed by Wang as a second transgene. An ordinarily skilled artisan would have been motivated to further include the Sox9 expressing plasmid as Wang teaches that Sox9 is a cartilage specific transcription factor and plays an essential role in chondrocyte differentiation and cartilage formation. Additionally, Wang demonstrates that Sox9 overexpression facilitates chondrogenesis with potential in cartilage tissue engineering. An ordinarily skilled artisan would have had a reasonable expectation of success as Glass, Sulaiman, and Wang are all drawn towards the use of MSCs and cartilage tissue repair/engineering.
Claims 12, 14, 22, and 35 are rejected under 35 U.S.C. 103 as being unpatentable over Glass, K.A., et al (2014) Tissue-engineered cartilage with inducible and tunable immunomodulatory properties Biomaterials 35; 5921-5931 in view of Morsut, L., et al (2016) Engineering customized cell sensing and response behaviors using synthetic Notch receptors Cell 164; 780-791 and Supplementary materials and Sulaiman, S.B., et al (2021) Type II collagen-conjugated mesenchymal stem cells micromass for articular tissue targeting Biomedicines 9(880); 1-17 as applied to claims 1, 11, and 32 above, and in further view of Kangari, P., et al (2020) Mesenchymal stem cells: amazing remedies for bone and cartilage defects Stem cell research & therapy 11(492); 1-21.
The combination of Glass, Morsut, and Sulaiman teach the methods of claim 11 and 32 as discussed in detail above.
The combination of applied references, however, does not explicitly disclose that the subject is in need of increased bone mass and/or volume as recited in claim 12 or that the engineered cell is administered to the subject systemically as recited in claim 14.
Kangari teaches that diseases of the skeletal system are extensively widespread in aged populations and are considered to be one of the main causes of disability and morbidity. The most common disorders of the skeletal system include intervertebral discs, osteoporosis, bone fractures, osteogenesis imperfecta, osteoarthritis, and rheumatoid arthritis. Among various therapeutic approaches for the treatment of these diseases, stem cell therapy seems to be more promising. Stem cells are introduced into tissues to repair, replace, and treat a defect with or without the addition of an external gene (page 1, paragraph bridging columns).
Kangari teaches that osteoarthritis is one of the most common arthritis related chronic disorders characterized by articular cartilage degeneration, thickening of subchondral bone and osteophyte formation (page 8, left column, paragraph 3). In recent years, cell therapy, especially with stem cells, is applied for the regeneration of OA damages. By virtue of their high proliferative capacity, chondrogenic differentiation capabilities, and immunosuppressive activities, MSC based therapies have demonstrated acceptable efficacy in cartilage repair in animal and clinical studies (page 8, right column, paragraph 2).
Kangari also teaches that MSCs are involved in the bone healing process because of their potential to increase osteoinduction and osteogenesis. These cells play crucial roles in bone repair and regeneration by several mechanisms including facilitating cell migration, homing, angiogenesis, response to inflammation, and differentiation (page 9, right column, paragraph 2).
Kangari also provides table 2, which discusses preclinical and clinical studies of MSCs for the treatment of skeletal diseases. The table teaches that in a human model of OA with 4 patients using BM-MSCs, positive changes in all patients were found with clear bone formation in osteonecrosis patients and cartilage regeneration in the OA patients.
Kangari further teaches that, in response to inflammation and high levels of pro-inflammatory factors, such as IFN-gamma, TNF-alpha, and IL-1β, MSCs are stimulated to start producing anti-inflammatory factors such as NO, IDO and anti-inflammatory cytokines and chemokines, which is followed by immunosuppression. Three days after bone fracture, transplanted MSCs are capable of limiting tissue injury by significant reduction in IL-6, TNF-alpha, and IL-1β levels and preventing the progression of fibrosis and thus improving bone regeneration (page 10, right column, paragraph 1). Kangari teaches that IL-1RA released by MSCs induces IL-10 in stimulated DCs and inhibits TNF-alpha production by activated macrophages which results in accelerated bone healing (page 11, right column, paragraph 1).
Kangari also discusses models in which MSC were administered by systemic injection (page 6, right column, paragraph 1).
It would have been prima facie obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to modify the method disclosed by the combination of Glass, Morsut, and Sulaiman by treating a patient who also is in need of increased bone mass or volume, such as a patient with osteonecrosis, as taught by Kangari, or to administer the engineered cells systemically as taught by Kangari and further supported by Sulaiman. It would have been obvious to treat a patient who is in need of increased bone mass or volume as Kangari teaches that, in patients with OA, MSC therapy resulted in clear bone formation in osteonecrosis patients and cartilage regeneration. Additionally, Kangari establishes that IL-1Ra expression results in accelerated bone healing, and the engineered cells taught by the combination of Glass, Morsut, and Sulaiman comprise a transgene for the overexpression of IL-1Ra.
It would have been obvious to administer the engineered cell systemically as Kangari teaches studies in which MSC have been injected systemically, thus providing a reasonable expectation of success. Additionally, an ordinarily skilled artisan would have had a reasonable expectation of success as the engineered cell taught by the combination of Glass, Morsut, and Sulaiman comprises a type II collagen antibody as the external domain of the receptor which Sulaiman teaches is a potential method for targeted delivery of MSCs in therapy of cartilage defects and osteoarthritis.
Regarding claims 22 and 35, Kangari further teaches that bone fracture can occur under the circumstances of continuous mechanical stress, trauma, and some diseases, such as osteoporosis and cancer. Healing of this type of injury is a complex, regenerative process with the involvement of numerous cell types including progenitor, inflammatory, endothelial, and hematopoietic cells as well as growth factors such as TGF-β (page 7, left column, paragraph 2).
It would have been prima facie obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to use the method disclosed by the combination of Glass, Morsut, and Sulaiman to treat a subject who has cancer, for instance a subject who has a bone fracture due to cancer, based on the teachings of Kangari. It would have been obvious to treat a subject with a bone fracture from cancer as Kangari teaches that IL-1RA released by MSCs induces IL-10 in stimulated DCs and inhibits TNF-alpha production by activated macrophages which results in accelerated bone healing and also teaches that growth factors, cytokines, and signaling molecules, including TGF-β stimulate the synthesis of collagen type II, and the engineered cells taught by the combination of Glass, Morsut, and Sulaiman are MSCs overexpressing IL-1RA through a synthetic receptor that targets collagen type II. Thus, an ordinarily skilled artisan would have had a reasonable expectation of success.
Claims 24, 31, and 41 are rejected under 35 U.S.C. 103 as being unpatentable over Glass, K.A., et al (2014) Tissue-engineered cartilage with inducible and tunable immunomodulatory properties Biomaterials 35; 5921-5931 in view of Morsut, L., et al (2016) Engineering customized cell sensing and response behaviors using synthetic Notch receptors Cell 164; 780-791 and Supplementary materials and Sulaiman, S.B., et al (2021) Type II collagen-conjugated mesenchymal stem cells micromass for articular tissue targeting Biomedicines 9(880); 1-17 as applied to claims 1, 11, 19, and 32 above, and in further view of Matas, J., et al (2019) Umbilical cord-derived mesenchymal stromal cells (MSCs) for knee osteoarthritis: repeated MSC dosing is superior to a single MSC dose and to hyaluronic acid in a controlled randomized phase I/II trial Stem Cells Translational Medicine 8; 215-224.
The combination of Glass, Morsut, and Sulaiman teach the methods of claim 11, 19, and 32 as discussed in detail above.
The combination of applied references, however, does not disclose that the administration comprises at least a second administration of the engineered cells as recited in claims 24 and 41 or that the subject is 60 years or older as recited in claim 31.
Matas teaches that knee osteoarthritis (OA) is a leading cause of pain and disability. Although conventional treatments show modest benefits, pilot and phase I/II clinical trials with MSCs point to the feasibility, safety, and occurrence of clinical and structural improvement in focal or diffuse disease. The study discussed by Matas aimed to assess the safety and efficacy of the intra-articular injection of a single or repeated umbilical cord derived MSCs in knee OA. Only MSC treated patients experienced significant pain and functional improvements from baseline. At 12 months, WOMAC-A pain subscale reached significantly lower levels of pain in the MSC-2 treated group (which had a repeated second dose at 6 months) compared to HA controls. Pain visual analog scale was also lower in the MSC-2 group as well as the total WOMAC scale. Matas teaches that repeated UC-MSC treatment is safe and superior to active comparator in knee OA at 1-year follow up (abstract).
Matas also teaches that the target population included individuals that were 40-65 years of age (page 216, left column, paragraph 5, patients).
It would have been prima facie obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to modify the method disclosed by the combination of Glass, Morsut, and Sulaiman to include at least a second administration of the engineered cells and/or to treat patients who are 40-65 years of age based on the teachings of Matas.
An ordinarily skilled artisan would have been motivated to provide a second administration as Matas teaches that repeated MSC dosing is superior to a single MSC dose and to hyaluronic acid controls in the treatment of OA (title). An ordinarily skilled artisan would have had a reasonable expectation of success as Matas is teaching the treatment of osteoarthritis with MSC, which is the same disease and cells used in the method taught by the combination of Glass, Morsut, and Sulaiman.
It would have been obvious to treat a subject who is 40-65 years of age as Matas demonstrates that this age range was the target population in clinical trials in which MSCs were used to treat OA. The age range taught by Matas overlaps with the instantly claimed age range of 60 years or older rendering the instant claim obvious per MPEP 2144.05.I as it would have been obvious to treat a subject who is 40-65 including those that overlap with the instantly claimed range. An ordinarily skilled artisan would have had a reasonable expectation of success as Matas is teaching the treatment of osteoarthritis with MSC, which is the same disease and cells used in the method taught by the combination of Glass, Morsut, and Sulaiman.
Claims 27 and 44 are rejected under 35 U.S.C. 103 as being unpatentable over Glass, K.A., et al (2014) Tissue-engineered cartilage with inducible and tunable immunomodulatory properties Biomaterials 35; 5921-5931 in view of Morsut, L., et al (2016) Engineering customized cell sensing and response behaviors using synthetic Notch receptors Cell 164; 780-791 and Supplementary materials and Sulaiman, S.B., et al (2021) Type II collagen-conjugated mesenchymal stem cells micromass for articular tissue targeting Biomedicines 9(880); 1-17 as applied to claims 1, 11, and 32 above, and in further view of Eckstein, F., et al (2005) Accuracy and precision of quantitative assessment of cartilage morphology by magnetic resonance imaging at 3.0T Arthritis & Rheumatism 52(10); 3132-3136.
The combination of Glass, Morsut, and Sulaiman teach the methods of claim 11, 19, and 32 as discussed in detail above.
The combination of applied references, however, does not explicitly disclose that the bone and/or cartilage mass and/or volume is assessed following administration of the engineered cell.
Eckstein teaches that quantitative magnetic resonance imaging (MRI) of articular cartilage represents a powerful tool in osteoarthritis (OA) research, but has so far been confined to a field strength of 1.5T. Eckstein studied the precision of quantitative MRI assessments of human cartilage morphology at 3.0T with validated measurements at 1.5T. Eckstein performed MR images of the knee in 15 participants with OA and 15 healthy control subjects cartilage volume, thickness, and surface rea of the femorotibial cartilage plates were quantified. Eckstein teaches that quantitative MRI measurements of cartilage morphology at 3.0T were found to be accurate and tended to be more reproducible than at 1.5T. Imaging at 3.0T may therefore provide superior ability to detect changes in cartilage status over time and to determine responses to treatment with structure modifying drugs (abstract, Objective, methods, results, and conclusion).
It would have been prima facie obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to modify the method taught by the combination of Glass, Morsut, and Sulaiman by further assessing the cartilage volume, thickness, and surface area using the MRI techniques disclosed by Eckstein after administration of the engineered cells. An ordinarily skilled artisan would have been motivate to assess cartilage volume of treatment in order to determine response to treatment caused by the structure modifying therapeutic as suggested by Eckstein. An ordinarily skilled artisan would have had a reasonable expectation of success as Eckstein is teaching techniques for monitoring cartilage changes after treatment in osteoarthritis, which is the same disease being treated by the combination of Glass, Morsut, and Sulaiman. Additionally, Eckstein teaches that MRI is a powerful tool in OA research suggesting that its use was common in OA evaluation.
Conclusion
No claims are allowed.
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/AUDREY L BUTTICE/Examiner, Art Unit 1647
/SCARLETT Y GOON/Supervisory Patent Examiner
Art Unit 1693
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