DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Election/Restrictions
Claims 17-19 and 32-33 have been withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 03/03/2025.
Claim Status
Claims 5, 7, 11, 16, 20-24, 26-27, 31, 34, and 36 are canceled, claims 17-19 and 32-33 are withdrawn, claims 1-4, 8, 17, 35, and 37 are amended, claims 40-41 are new, and claims 1-4, 6, 8-10, 12-15, 25, 28-30, 35, and 37-41 have been considered on their merits.
It is noted that Applicant filed a supplemental amendment on 01/05/2026 which included a copy of the amended claim set from 11/21/2025 and a corrected argument section.
Withdrawn Rejections/Objections
The claim rejections under 35 U.S.C. § 103 have been withdrawn due to the instant amendments, however, new rejections are set forth below.
Claim Rejections - 35 USC § 103
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 (i.e., changing from AIA to pre-AIA ) 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.
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, 35, 37, and 40-41 are rejected under 35 U.S.C. 103 as being unpatentable over Studer et al. (WO 2018/175574, published 27 September 2018, of record) in view of George et al. (US 2018/0021383, of record).
This is a new rejection, necessitated by applicant’s amendments to the claims. A response to applicant' s traversal follows the new rejection below.
Regarding claim 1 and 6, Studer et al. teach in vitro methods for inducing differentiation of stem cells, wherein the stem cells are human iPSCs (hiPSCs) (p. 28, Section 5.2, lines 19-23). Studer et al. teach a method for derivation of anterior forebrain neuroectoderm from hPSCs wherein hPSC were dissociated into single cells and plated onto Matrigel coated dishes in stem cell maintenance media containing ROCK inhibitor (Y-27632) (p. 65, lines 17-20). Studer et al. teach the media was changed the next day to a neural induction media (p. 65, lines 20-21), this reads as the hPSCs were exposed to a RHO kinase inhibitor for at least 8 hours up to 3 days, as the next day reads as approximately 24 hours which falls in the necessary range.
Studer et al. is silent to the method in which the monolayer of cultured iPSCs was generated. However, mechanical separation (shaking) of cells is well-known in the art. George et al. teach PSCs are dissociated into essentially individual cells using mechanical or enzymatic methods known in the art (George, para. [0126]). Therefore, it would have been obvious to utilize the method of George et al. to mechanically separate the iPSCs of Studer et al. with a reasonable expectation of success because George et al. teach methods of mechanically dissociating cells is known in the art.
Studer et al. teach the next day, the media was changed to a neural induction media (a first media) comprising LDN193189 and SB431542 which represents day 0 of differentiation (p. 65, lines 20-23). Studer et al. teach between day 10-12 of differentiation, cells were either used to generate rosette-stage NSCs or long-term human embryonic stem cell-derived neural stem cells (LTNSCs) (p. 65, lines 26-27). Day 10-12 of differentiation reads as the cells were cultured in the first media for 7-13 days, as 10-12 fall within the range of 7-13 days.
Studer et al. teach a method of generating LTNSCs wherein differentiated cells were dissociated and separated into clumps (aggregates) and resuspended in N2 media containing FGF wherein the cells should form a high number of neurospheres (claim 6) (p. 66, lines 4-7). Studer et al. teach once the neurosphere cultures are pure the neurospheres are cultured on PO/Lam/FN plates in N2 with FGF2 and EGF (p. 66, lines 7-9).
Studer et al. teach regionally patterned neural stem cells are infected with lentiviral particles containing FUW-NFIA and FUW-M2-rtTA and induced with doxycycline (dox) one day after infection for 5 days then switched to the astrocyte induction media (N2) with HB-EGF and LIF without dox for a minimum of 5 days (p. 66, lines 20-25). Studer et al. teach, in certain embodiments, the regionally patterned progenitors are cortical progenitors (p. 30, lines 16-17). Regionally patterned neural stem cells which are cortical progenitors reads as cortical neuronal progenitor cells (NPCs).
Studer et al. teach the in vitro method for inducing differentiation of stem cells into astrocytes and precursors thereof comprises exposing a population of stem cells with effective amounts of (i) one or more inhibitor of transforming growth factor beta (TGFβ)/Activin-Nodal signaling and/or one or more inhibitor of bone morphogenetic protein (BMP) signaling, (ii) one or more NFIA activator (EGF and FGF), and (iii) LIF (p. 46, lines 5-9). The second and third steps read as the second media comprises EGF, FGF, and LIF.
Studer et al. teach the neurospheres in the second media are cultured on plates and is silent to generating iPSC-derived cortical NPCs in suspension culture of the second media, specifically.
However, George et al. teach differentiation of iPSCs can be induced by leaving the cells in suspension in the presence of a proliferation-inducing growth factor, without dissociating the neurospheres (para. [0128]). Therefore, it would have been obvious to one of ordinary skill in the art to generate the iPSC-derived cortical NPCs in the second media of Studer et al. in a suspension culture as taught by George et al. with a reasonable expectation of success because George et al. teach differentiation of the iPSCs can also be induced by leaving the cells in suspension in the presence of a proliferation-inducing growth factor, without dissociating the neurospheres. One would be motivated to generate the iPSC-derived cortical NPCs in the second media of Studer et al. in a suspension culture as taught by George et al. because one of the intermediate steps of Studer et al. involves forming aggregated neurospheres and George et al. teach this step can be successfully accomplished in suspension.
Regarding the wherein clause in the last two lines of claim 1, wherein the iPSC-derived NPCs are capable of expanding as spheres or aggregates and differentiating into astrocytes. The wherein clause, “wherein the iPSC-derived NPCs are capable of expanding as spheres or aggregates”, does not limit the claimed method as it is not directed to an active step and merely stating the characteristics of the iPSC-derived cortical NPCs. Therefore, if the references teach the active steps of the method then any the iPSC-derived cortical NPCs derived from the same method would be expected to have the same characteristics.
However, if this clause was limiting, Studer et al. teach a method of generating LTNSCs wherein differentiated cells were dissociated and separated into clumps (aggregates) and resuspended in N2 media containing FGF wherein the cells should form a high number of neurospheres (p. 66, lines 4-7).
Regarding claim 35, Studer et al. teach stem-cell-derived glial competent cells (e.g., astrocyte precursors) and astrocytes, the method comprising in vitro differentiation of stem cells to NSCs, in vitro differentiation of NSCs to glial competent cells, and in vitro differentiation of glial competent cells to astrocytes (p. 29, lines 3-7 and p. 45, lines 16-35). Regarding the markers of cortical neural progenitors, Studer et al. teach LIF was most efficient in generating GFAP+ cells (p. 71, lines 26-29). Studer et al. teach an in vitro method for differentiating pluripotent stem cells, comprising promoting NFIA signaling in a population of cells expressing one or more neural stem cell marker to obtain a cell population comprising at least about 10% differentiated cells and expressing one or more glial competent cell marker and wherein the cell population comprise one or more astrocyte markers GFAP, AQP4, CD44, S100β, SOX9, NFIA, GLT-1, and CSRP1 (claims 1 and 35). Therefore, Studer et al. teach cells with markers for both cortical neural progenitors and markers of genes associated with both mature astrocytes and immature astrocytes, as the instant specification at para. [0090] disclose neural progenitor markers include GFAP and S100β and discloses markers for astrocytes include GFAP, S100β, and GLT-1.
Regarding claim 37, wherein the iPSC-derived NPCs are capable of developing filopodia in culture and expressing neural progenitor markers including Nestin. The wherein clause, “wherein the iPSC-derived NPCs are capable of developing filopodia in culture”, does not limit the claimed method as it is not directed to an active step and merely stating the characteristics of the iPSC-derived NPCs. Any NPCs would be considered to have the claimed feature, absent evidence to the contrary. Additionally, Studer et al. teach the neural stem cell marker to include NESTIN (claim 21).
Regarding claim 40, Studer et al. teach derivation of anterior forebrain neuroectoderm from hPSCs were dissociated into single cells and plated onto Matrigel (BD Biosciences) coated dishes in stem cell maintenance media containing 10 µM ROCK inhibitor (Y-27632) (p. 65, lines 17-20). This concentration falls in the range of about 0.5 µM to about 12 µM.
Regarding claim 41, Studer et al. teach stem cells are contacted with one or more BMP inhibitor (LDN) in a concentration of about 10 nM to about 500 nM (p. 50, lines 33-34). 10 nM LDN193189 is approximately 4.1 ng/mL and 500 nM is approximately 0.24 µg/mL. Studer et al. teach stem cells are contacted with one or more inhibitor of TGFβ/Activin-Nodal signaling (SB) in a concentration of about 1 µM to about 20 µM (p. 50, lines 20-21). Studer et al. teach stem cells are contacted with one or more FGF activator in a concentration from about 1 ng/ml to 100 ng/ml (p. 40 lines 13-15). Studer et al. teach stem cells are contacted with one or more EGF-family protein in a concentration from about 1 ng/ml to 100 ng/ml (p. 40, lines 33-34). Studer et al. teach glial competent cells are contacted by an effective amount of LIF to increase detectable levels of one or more astrocyte markers (p. 45, lines 27-29). Studer et al. teach for astrocyte induction cells were contacted by 10 ng/ml LIF for a minimum of 5 days (p. 66, lines 24-25).
Regarding the concentration of SB, FGF, and EGF, where the claimed ranges overlap or lie inside ranges disclosed by the prior art a prima facie case of obviousness exists: selecting the specific concentration, is obvious (see M.P.E.P. § 2144.05). One would have had a reasonable expectation of successfully selecting concentration of small molecules and growth factors which falls inside the concentrations taught by Studer et al. due to the variables of the conditions of the culturing environment, cell source variation, and density of the cells being differentiated.
Additionally, M.P.E.P. § 2144.05(II)(A) states a prima facie case of obviousness exists "[W]here the general conditions of a claim are disclosed in the prior art, it is not inventive to discover the optimum or workable ranges by routine experimentation." In re Aller, 220 F.2d 454, 456, 105 USPQ 233, 235 (CCPA 1955). Furthermore, the concentrations of LDN and LIF would have been a result effective variable amenable to routine optimization and experimentation on the part of the artisan of ordinary skill, said artisan recognizing the concentration required to enhance efficiency, depends on the density of cells, and cell source variation. A holding of obviousness over the cited claims is therefore clearly required. The normal desire of scientists or artisans to improve upon what is already generally known provides the motivation to determine where in a disclosed set of concentration ranges is the optimum combination of concentrations. See Peterson, 315 F.3d at 1330, 65 USPQ2d at 1382.; See also M.P.E.P. § 2144.05 (II)(A).
Therefore, the invention as a whole would have been prima facie obvious to a person of ordinary skill before the effective filing date of the claimed invention.
Response to Traversal
Applicant's arguments filed 05 January 2026 have been fully considered but they are not persuasive.
Applicant argues on page 8 of the response, nowhere in Studer does it disclose or suggest that anterior forebrain neuroectoderm comprises cortical NPC and these terms are not interchangeable.
This is not found persuasive because Studer does teach generating cortical NPCs. Specifically, Studer et al. teach, in certain embodiments, the regionally patterned progenitors are cortical progenitors (p. 30, lines 16-17). Regionally patterned neural stem cells which are cortical progenitors reads as cortical neuronal progenitor cells (NPCs). As such, the rejections have been revised to address the newly amended claims. Therefore, the arguments set forth are not persuasive.
Applicant argues Studer does not teach cortical NPCs, yet points out on page 8 of the response, Studer teach hESC derived neural stem cells which resemble early neuroectoderm. The neuroectoderm is known to give rise to the entire nervous system, which would include cortical NPCs. Applicant also highlights Studer teach cortical neurons were derived by differentiating hPSCs towards the neuroectodermal fate, if Studer teach cortical neurons derived from hPSCs, then cortical NPCs would have necessarily resulted from the methods of Studer. Therefore, the arguments set forth are not persuasive.
Applicant argues on page 9 of the response, the method of Studer at page 65 relates to derivation of anterior forebrain neuroectoderm from hPSCs, however, it is noted Studer teaches the cells were used to generate LTNSCs. Studer teaches LTNSCs express several neural stem cell markers and neural stem cells are considered progenitor cells. Therefore, the arguments set forth are not persuasive.
Applicant argues on page 10 of the response, the neural induction media of Studer also comprises KSR in addition to LDN and SB. In response to applicant's argument that the references fail to show certain features of the invention, it is noted that the features upon which applicant relies (i.e., the media not comprising KSR) are not recited in the rejected claim(s). Although the claims are interpreted in light of the specification, limitations from the specification are not read into the claims. See In re Van Geuns, 988 F.2d 1181, 26 USPQ2d 1057 (Fed. Cir. 1993). The language of the claim does not limit additional media components.
Applicant argues on page 11 of the response, the neural induction media of Studer also comprises XAV939. In response to applicant's argument that the references fail to show certain features of the invention, it is noted that the features upon which applicant relies (i.e., the media not comprising XAV939) are not recited in the rejected claim(s). Although the claims are interpreted in light of the specification, limitations from the specification are not read into the claims. See In re Van Geuns, 988 F.2d 1181, 26 USPQ2d 1057 (Fed. Cir. 1993). The language of the claim does not limit additional media components.
Applicant argues on page 13 of the response, Studer in view of George would not “to arrive at a "method to generate induced pluripotent stem cell (iPSC)-derived cortical neuronal progenitor cells (NPCs)," "wherein the iPSC-derived cortical NPCs are capable of expanding as spheres or aggregates and differentiating into astrocytes," as recited in amended claim 1.” This statement does not limit the claimed method as it is not directed to an active step and merely stating the characteristics of the iPSC-derived cortical NPCs. Therefore, if the references teach the active steps of the method then any the iPSC-derived cortical NPCs derived from the same method would be expected to have the same characteristics.
Claim 2 is rejected under 35 U.S.C. 103 as being unpatentable over Studer et al. (WO 2018/175574, published 27 September 2018, of record) in view of George et al. (US 2018/0021383, of record) as applied to claims 1, 6, 35, 37, and 40-41 above, and further in view of Efthymiou et al. (Expert Opin Biol Ther. 2014 September; 14(9)).
This is a new rejection, necessitated by applicant’s amendments to the claims.
Regarding claim 2, the claim language does not limit the order of steps, nor require the cells to be cultured in the presence of the ROCK inhibitor while in suspension. Studer et al. teach a method for derivation of anterior forebrain neuroectoderm from hPSCs wherein hPSC were dissociated into single cells (reads as provided in suspension) and plated onto Matrigel coated dishes in stem cell maintenance media containing ROCK inhibitor (Y-27632) (reads as cultured in the presence of the RHO kinase inhibitor) (p. 65, lines 17-20). Studer et al. teach the media was changed the next day to a neural induction media (p. 65, lines 20-21). This step in the method of Studer et al. reads as the cells were cultured in the presence of the RHO kinase inhibitor prior to being plated and treated with the first media because the media was changed and the ROCK inhibitor was not present in the first media. Therefore, the RHO kinase inhibitor was administered prior to treatment with the first media.
Even if the limitations of the claim required the stem cells to be cultured in the presence of a ROCK inhibitor while in suspension, Efthymiou et al. teach cell-to-cell contact is essential for hPSC survival; hPSCs generally do not survive dissociation by trypsin/EDTA and instead are dissociated with the enzymes collagenase and dispase, which generate clumps of cells that preserve cell-to-cell contact (p. 3, Section 3.3). Efthymiou et al. teach it is difficult to regulate the size of these clumps, which results in cell culture variability (p. 3, Section 3.3). Efthymiou et al. teach these animal-sourced enzymes, additionally, are not ideal for translational research (p. 3, Section 3.3). Efthymiou et al. teach the addition of ROCK inhibitor has been shown to suppress cell death, thereby allowing for dissociation methods with more consistent cell survival in regular expansion and differentiation (p. 3, Section 3.3). Therefore, it would have been obvious to culture the dissociated PSCs in of Studer et al. with the ROCK inhibitor while in suspension with a reasonable expectation of success because both references teach culturing PSCs in the presence of a ROCK inhibitor and Efthymiou et al. teach advantages in doing so while the PSCs are in suspension. One would be motivated to culture the dissociated PSCs in of Studer et al. with the ROCK inhibitor while in suspension because Efthymiou et al. teach ROCK inhibitors suppress cell death in dissociated cells in culture.
Therefore, the invention as a whole would have been prima facie obvious to a person of ordinary skill before the effective filing date of the claimed invention.
Claim 3 is rejected under 35 U.S.C. 103 as being unpatentable over Studer et al. (WO 2018/175574, published 27 September 2018, of record) in view of George et al. (US 2018/0021383, of record) as applied to claims 1, 6, 35, 37, and 40-41 above, and further in view of Clementi et al. (Cell Engineering and Regeneration, Springer Nature 2018).
This is a new rejection, necessitated by applicant’s amendments to the claims. A response to applicant' s traversal follows the new rejection below.
Regarding claim 3, Studer et al. teach the cultured iPSCs were cultured in a ROCK inhibitor for a day, as discussed in the rejection of claim 1. Studer et al. is silent to the method in which the monolayer of cultured iPSCs was generated.
However, Clementi et al. teach the advantages of stem cell expansion and differentiation under dynamic conditions (Abstract). Clementi et al. teach intentional active motion enhances mass transfer and mechanotrasductive effects which often results in higher numbers of functional cells (Abstract). Clementi et al. teach orbital (X-Y motion) shaker, are known in the art (p. 6).
Regarding the effect of X-Y motion as it relates to even distribution of cells, this would be a matter of routine optimization on the part of the artisan of ordinary skill, said artisan recognizing the shaker parameters in order to achieve the desired results. A holding of obviousness over the cited claims is therefore clearly required. The normal desire of scientists or artisans to improve upon what is already generally known provides the motivation to determine where in a disclosed set of percentage ranges is the optimum combination of percentages. See Peterson, 315 F.3d at 1330, 65 USPQ2d at 1382.; See also M.P.E.P. § 2144.05 (II)(A).
Therefore, the invention as a whole would have been prima facie obvious to a person of ordinary skill before the effective filing date of the claimed invention.
Response to Traversal
Applicant’s arguments on page 10 of the response, with respect to the rejection of claim 3 under 103 have been fully considered and are persuasive. The previously cited art does not teach the newly amended claim. Therefore, the rejection has been withdrawn. However, upon further consideration, a new ground(s) of rejection is made in view of Clementi et al.
Claims 4 and 38-39 are rejected under 35 U.S.C. 103 as being unpatentable over Studer et al. (WO 2018/175574, published 27 September 2018, of record) in view of George et al. (US 2018/0021383, of record), as applied to claims 1, 6, 35, 37, and 40-41 above, and in view of Fernandopulle et al. (Current Protocols in Cell Biology, 2018, of record).
This is a new rejection, necessitated by applicant’s amendments to the claims.
Regarding claims 4 and 38, Studer et al. teach between day 10-12 of differentiation in the first media, cells were either used to generate rosette-stage NSCs or LTNSCs (p. 65, lines 26-27).
Studer et al. is silent to cells scraped from the monolayer culture in the first media specifically, however, scraping iPSCs is well known in the art.
Fernandopulle et al. teach pipette tips or specialized cell scraper can be used depending on the scale of cells to be removed (p. 12, pick-to-remove). Fernandopulle et al. teach iPSCs can be gently scraped until cells are in suspension (p. 12, step 46b). Therefore, it would have been obvious to one of ordinary skill in the art to scrape the cells from the monolayer culture to be cultured in the second media of Studer et al. with a reasonable expectation of success because scraping iPSCs to put the cells in suspension is a well-known method in the art. One would be motivated to scrape the cells from the monolayer culture of Studer et al. because it is a known technique for removing cells from an adhesion culture as indicated by Fernandopulle et al.
Regarding claim 39, Studer et al. teach regionally patterned neural stem cells are infected with lentiviral particles containing FUW-NFIA and FUW-M2-rtTA and induced with doxycycline (dox) one day after infection for 5 days then switched to the astrocyte induction media (N2) with HB-EGF and LIF without dox for a minimum of 5 days (p. 66, lines 20-25). Switching to the N2 media reads as the second media is absent of LDN and SB.
Therefore, the invention as a whole would have been prima facie obvious to a person of ordinary skill before the effective filing date of the claimed invention.
Claims 8-9 are rejected under 35 U.S.C. 103 as being unpatentable over Studer et al. (WO 2018/175574, published 27 September 2018, of record) in view of George et al. (US 2018/0021383, of record), as applied to claims 1, 6, 35, 37, and 40-41 above, and further in view of Zhang et al. (BioMed Research International, 2015, of record).
This is a new rejection, necessitated by applicant’s amendments to the claims.
Regarding claim 8, Studer et al. teach LTNSCs express NSC markers NESTIN (p. 4, line 31 and Fig 1C), βIII-Tubulin (TUBB3) (Fig 1F) , and quantitate PCR analysis showed the presence of S100β in glial competent expression (pp. 20/21, lines 30-33/lines 1-2 and Fig 21B and C). Regarding the limitation directed to introducing at least one vector into the iPSCs-derived cortical NPCs, Studer et al. teach regionally patterned neural stem cells are infected with lentiviral particles (at least one vector) containing FUW-NFIA and FUW-M2-rtTA and induced with doxycycline (dox) one day after infection for 5 days then switched to the astrocyte induction media (N2) with HB-EGF and LIF without dox for a minimum of 5 days (p. 66, lines 20-25).
Studer et al. is silent to the cell marker VIM (Vimentin), although it is well known in the art Nestin and VIM are often co-expressed in neural progenitor cells.
Additionally, Zhang et al. teach Nestin is a neuroepithelial stem cell protein which is expressed in immature neural progenitor cells (p. 7, section 3.2.13), Vimentin (VIM) is an intermediate filament protein which imply the property of glial cells and is exhibited in immature astrocytes (p. 7, section 3.2.20), Tuj1, which is synonymous with TUBB3, is a neuron-specific class III β-tubulin and can be detected in immature neurons (p. 8, section 3.2.25), and S100β can be detected of specific postmitotic astrocytes and indicates a more mature stage (p. 8, section 3.2.21). Therefore, these cell markers are known neural progenitor cell/astrocytes markers, thus, it would have been obvious to one of ordinary skill in the art to have iPSC-derived cortical NPCs with the indicated cell markers if one cultured iPSCs in a media directed toward producing astrocytes. There would have been a reasonable expectation of success in expressing these specific cell markers because iPSC-derived cortical NPCs are capable of differentiating into astrocytes, absent evidence to the contrary.
Regarding claim 9, Studer et al. teach non-viral approaches can also be employed for delivering the NFIA nucleic acid to the cells to include transfection in vitro using calcium phosphate or electroporation (pp. 37/38 lines 30-34/1).
Therefore, the invention as a whole would have been prima facie obvious to a person of ordinary skill before the effective filing date of the claimed invention.
Claims 10, 12-13, 25, and 28-30 are rejected under 35 U.S.C. 103 as being unpatentable over Studer et al. (WO 2018/175574, published 27 September 2018) in view of George et al. (US 2018/0021383, of record) and Zhang et al. (BioMed Research International, 2015, of record), as applied to claims 8-9 above, and further in view of Akhtar et al. (Stem Cell Reports, of record).
This is a new rejection, necessitated by applicant’s amendments to the claims.
Regarding claim 10 and 12-13, Studer et al. teach expression of the NFIA nucleic acid is operably linked to an inducible promoter (p. 37, lines 9-10). This reads as an expression cassette comprising a constitutive or inducible promoter operably linked to a sequence encoding a protein or peptide.
Studer et al. is silent to at least one homologous recombination sequence; or
at least one vector comprises a piggyBac transposon-based vector and a pBase vector, and the piggyBac transposon-based vector comprises: an expression cassette, comprising: a constitutive promoter; an inducible, bi-directional polycistronic promoter comprising a tet responsive element; and a sequence encoding a protein or peptide; two transposon elements, wherein the two transposon elements flank the expression cassette; and at least one homologous recombination sequence.
However, Akhtar et al. teach incorporating constitutive expression of the rtTA-V10 transactivator and a dox-responsive TRE-Bi (Tet response element bidirectional) promoter, as well as reporter genes in a single system requires a large vector, which was accomplished with a vector named ‘‘pB-RTP Tet- GDNF/memClover-FLuc’’ (piggyBac-reverse transactivator/TagBFP2nls/PacR-Tet-inducible-GDNF/membrane Clover-firefly luciferase) (p. 1697, Results 1st column and Figure 1A). The vector is flanked by piggyBac terminal repeats (PB TR) (reads as two transposon elements flank the expression cassette) (Figure 1A1), which allows for its stable genomic integration when the pBase enzyme is transiently expressed (p. 1697, Results 1st column). The expression of the pBase enzyme reads as the vector comprises a pBase vector, also see Figure 1B. Akhtar et al. teach the vector has two promoters, a constitutively active CMV/Chick b-actin (also known as CAG) promoter and an inducible, bidirectional TRE-Bi promoter (Tet response element bidirectional) (p. 1697, Results 1st column). Akhtar et al. teach the CAG promoter (Figure 1A2) drives constitutive expression of the rtTA-V10 (also known as tet-ON) transactivator (Figure 1A3), TagBFP2-V5-nls (enhanced blue fluorescent protein with a V5 tag and nuclear localization sequence) (p. 1697, Results 1st column). The TagBFP2-V5-nls reads as a sequence encoding a protein. The glial cell line-derived neurotrophic factor (GDNF) (claims 12-13) of the above-named vector reads as a homologous recombination sequence. The constitutive promoter, promoter comprising a tet responsive element, a sequence encoding protein, two transposon elements, and the homologous recombination sequence read as an expression cassette.
Therefore, it would have been obvious to one of ordinary skill in the art to utilize the vector of Akhtar et al. in the method of Studer et al. with a reasonable expectation of success because Akhtar et al. teach human neural progenitor cells genetically engineered to stably produce glial cell line derived neurotrophic factor (GDNF) can survive, migrate, release GDNF, and protect degenerating neurons (Akhtar, p. 1696, 1st para.). One would have been motivated to utilize the vector of Akhtar et al. in the method of Studer et al. because Akhtar et al. teach combined cell and gene therapy approaches to both rejuvenate cellular niches and provide therapeutic molecules to diseased host cells is a promising treatment approach for neurological disorders (Akhtar, p. 1696, 1st para.).
Regarding claim 25, Akhtar et al. teach GDNF transcription with previous lentiviral experiments is dependent on the PGK promoter that is activated by endogenous transcription factors (p. 1702, 1st column). Therefore, the PGK promoter was known in the art to promote GDNF transcription.
Regarding claims 28 and 29 directed to the inducible promoter operably linked to the sequence encoding the protein comprising a promoter regulated by a tetracycline-class antibiotic/doxycycline, Akhtar et al. teach a doxycycline-regulated vector, allowing inducible and reversible expression of a therapeutic molecule (Abstract). Akhtar et al. teach doxycycline regulates GDNF expression from pB-RTP-Tet-GDNF/memClover-FLuc nucleofected iPSC-derived NPCs (Figure 2).
Regarding claim 30 directed to the inducible promoter operably linked to the sequence encoding the protein comprising a promoter is regulated by a reverse tetracycline-controlled transactivator (rtTA), Akhtar et al. teach the vector has two promoters; a CAG promoter and an inducible, bidirectional TRE-Bi promoter (p. 1697 Results 1st column). The CAG promoter (Figure 1A2) drives constitutive expression of the rtTA-V10 (also known as tet-ON) transactivator (Figure 1A3) (p. 1697 Results 1st column). The rtTA-V10 reads as the sequence encoding the protein comprising a promoter is regulated by an rtTA.
Therefore, the invention as a whole would have been prima facie obvious to a person of ordinary skill before the effective filing date of the claimed invention.
Claims 14-15 are rejected under 35 U.S.C. 103 as being unpatentable over Studer et al. (WO 2018/175574, published 27 September 2018) in view of George et al. (US 2018/0021383, of record) and Zhang et al. (BioMed Research International, 2015, of record) and Akhtar et al. (Stem Cell Reports, of record), as applied to claims 10, 12-13, 25, and 28-30 above, and further in view of Oceguera-Yanez et al. (Methods, Volume 101, of record).
This is a new rejection, necessitated by applicant’s amendments to the claims.
Regarding claims 14 and 15 directed to a homologous recombination sequence capable of targeting a genomic safe harbor (AAVS1), Studer et al. in view of George et al., Zhang et al., and Akhtar et al. are silent to the limitation.
However, Oceguera-Yanez et al. teach the AAVS1 is an exemplary locus within the PPP1R12C gene that permits robust expression of CAG promoter-driven transgenes in iPSCs (Abstract). The transgenes read as a homologous recombination sequence. Oceguera-Yanez et al. teach gene expression, from the AAVS1 locus is maintained during long-term human iPSC culture and in vitro differentiation along multiple lineages (Abstract).
It would have been obvious to a person skilled in the art to use the method of Oceguera-Yanez et al. to insert a homologous recombination sequence capable of targeting a genomic safe harbor in the method of Studer et al. in view of George et al., Zhang et al., and Akhtar et al. with a reasonable expectation of success because the combined effect of these teachings would have been expected to be greater than the effect of either individually, as each aim to provide long-term stable expression of a homologous recombination sequence. A person of ordinary skill in the art would have been motivated to do so because Akhtar et al. teach targeting the AAVS1 safe landing may allow for long-term stable integration and methylation-resistant expression of transgenes (p. 1702, Discussion 2nd column). Transgenes read as a homologous recombination sequence. Therefore, the combined teachings of Studer et al. in view of George et al., Zhang et al., and Akhtar et al. and Oceguera-Yanez et al. teach the limitations of claims 14 and 15.
Therefore, the invention as a whole would have been prima facie obvious to a person of ordinary skill before the effective filing date of the claimed invention.
Conclusion
No claims are allowed.
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
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/N.A.H./Examiner, Art Unit 1631
/LAURA SCHUBERG/Primary Examiner, Art Unit 1631