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 .
Claims 4-14, 18, 25-28, 31, 33-35, 41, 44-52, 54, 55 have been canceled. Claims 1-3, 15-17, 19-24, 29, 30, 32, 36-40, 42, 43, 53, 56 remain pending.
Applicant's arguments filed 8-13-26 have been fully considered but they are not persuasive.
The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
When referencing the specification, please use page and paragraph number (and line number as necessary) of the original specification. Do not simply use paragraph numbers. Do not refer to the US Patent Application Publication. There are often discrepancies between the paragraph numbers of the original specification and the Published Application.
Election/Restrictions
Applicants elected Group I, claims 1-14, 18, 25-35, 40-45, 48, without traverse in the response filed 7-13-22. Claim 49 was combined with Group I in the office action sent 1-18-24 because it is equivalent to the cells of claims 44 and 45.
Claims 15-17, 19-24, 36-39 remain withdrawn.
Claims 1-3, 29, 30, 32, 40, 42, 43, 53, 56 remain under consideration.
Claim objections
Step A) of claim 1 can be written more clearly as ---i) culturing isolated mammalian neural stem cells that comprise a nucleic acid sequence encoding exogenous NFIA operably linked to an inducible promoter in the presence of an activator of the promoter such that glial cells that express CD44 are obtained---.
The phrase “without inducing expression of the exogenous NFIA” in step B) of claim 1 is awkward. It is missing the fact that the cells are cultured in the absence of the activator of the inducible promoter. Step B) can be written more clearly as ---ii) culturing the plurality of cells obtained in step i) in the presence of [LIF] for at least 3 days in the absence of the activator of the inducible promoter---.
Claim 2 can be written more simply as ---further comprising culturing mammalian pluripotent cells in the presence of SB431542, LDN193189, or a derivative thereof such that the mammalian neural stem cells are obtained---.
Claim 3 can be written more accurately as ---wherein step i) occurs at least 8 days after culturing the mammalian pluripotent cells in the presence of SB431542, LDN193189, or a derivative thereof such that the mammalian neural stem cells are obtained---.
Claim 29 can be written more simply as ---wherein step ii) occurs at least 10 days after culturing the mammalian pluripotent cells in the presence of SB431542, LDN193189, or a derivative thereof such that the mammalian neural stem cells are obtained---.
Claim Rejections - 35 USC § 112
Indefiniteness
Claims 1-3, 15-17, 19-24, 29, 30, 32, 36-40, 42, 43, 53, 56 remain 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.
Withdrawn rejections
The rejection regarding the metes and bounds of “glial competent neural stem cell that express CD44 or AQP4” as required in step A) of claims 1, 53, 55 or “glial competent cell that express GFAP” as required in step B) of claims 1, 53, 55 has been withdrawn because “glial competent” and “competent”, respectively, have been deleted.
The concept of “glial cells” that express GFAP and are “astrocyte precursors” in claim 32 is indefinite. While glial cells may be astrocytes and astrocytes may express GFAP, it is unclear how a glial cell that expresses GFAP can be an “astrocyte precursor”. The art taught that astrocyte precursors “Often express proteins like Vimentin or Pax2, while typically lacking mature astrocytic markers like GFAP”. Therefore, the glial cell expressing GFAP in claim 1 cannot be an “astrocyte precursor” as required in claim 32. This makes the claim indefinite.
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Written description
Claims 1-3, 15-17, 19-24, 29, 30, 32, 36-40, 42, 43, 56 remain rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention.
Withdrawn rejection
The rejection of claim 53 has been withdrawn because the claim has been limited to culturing mammalian neural stem cells in the presence of TGFβ1 for at least 5 days such that glial cells that express CD44 or AQP4 are obtained.
The rejection of claim 55 has been withdrawn because the claim has been canceled.
Pending rejection
A) The specification lacks written description for any means of inducing expression of exogenous NFIA in isolated mammalian neural stem cells that comprise a nucleic acid sequence encoding exogenous NFIA operably linked to an inducible promoter such that glial cells that express CD44 or AQP4 are obtained as broadly encompassed by claim 1 other than culturing isolated mammalian neural stem cells that comprise a nucleic acid sequence encoding exogenous NFIA operably linked to an inducible promoter in the presence of an activator of the promoter such that glial cells that express CD44 or AQP4 are obtained.
Claim 1 is drawn to:
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Claim 53 is drawn to:
Claim 1 as newly amended requires inducing expression of exogenous NFIA in neural stem cell that contain a nucleic acid sequence encoding an exogenous NFIA operably linked to an inducible promoter. Claim 1 as newly amended encompasses inducing expression of any exogenous NFIA expression by any means, e.g. TGFβ. It does not require expression of the exogenous NFIA in the exogenous nucleic acid sequence. It does not require culturing the cells in the presence of an activator of the inducible promoter.
However, the specification is limited to 1) culturing mammalian neural stem cells comprising an exogenous nucleic acid sequence encoding an exogenous NFIA operably linked to an inducible promoter in the presence of an activator of the promoter such that glial cells expressing CD44 or AQP4 are obtained (pg 59, lines 27-32) such that glial cells expressing CD44 or AQP4 are obtained (pg 71, lines 13-21). The specification does not correlate culturing the cells in the presence of an activator of the inducible promoter on pg 59 to the absence of an activator or to using any other means as broadly as encompassed by claim 1 as newly amended.
Accordingly, the claim lacks written description for inducing expression of exogenous NFIA in isolated mammalian neural stem cells that comprise a nucleic acid sequence encoding exogenous NFIA operably linked to an inducible promoter such that glial cells that express CD44 or AQP4 are obtained as broadly encompassed by claim 1 other than culturing isolated mammalian neural stem cells that comprise a nucleic acid sequence encoding exogenous NFIA operably linked to an inducible promoter in the presence of an activator of the promoter such that glial cells that express CD44 or AQP4 are obtained.
Response to argument
Applicants argue amendment overcomes the rejection. Applicants’ argument is not persuasive for reasons set forth above.
B) The concept of obtaining a population of cells comprising at least 10% neural stem cells that express CD44 in claims 1, 53, 55 lacks written description. CD44 is “expressed in a large number of mammalian cell types” (Wikipedia definition of CD44, “Tissue distribution and isoforms”, 2024). CD44 is not a marker of glial cells or some [nondisclosed] secondary neural stem cell (obtained by differentiating neural stem cells expressing i) Pax6, Nestin, Sox1, Sox2, Plzf, ZO1, and BRN2; or ii) Pax6, Sox1, Plzf, and ZO1).
Fig. 2 and 3 show CD44 (and AQP4, Sox2 and Nestin) expression but does not teach the cells that express CD44 (or AQP4, Sox2, or Nestin) are neural stem cells as required in claims 1, 53.
An adequate written description of a method requires more than a mere statement that it is part of the invention and reference to a potential method for isolating it; what is required is using CD44 or AQP4 expression as a marker of glial cells or describing the secondary neural stem cell obtained by differentiating neural stem cells expressing i) Pax6, Nestin, Sox1, Sox2, Plzf, ZO1, and BRN2; or ii) Pax6, Sox1, Plzf, and ZO1.
Accordingly, the specification lacks written description for obtaining 10% cells that express CD44 or AQP4, using them as “glial cells”, or as some [nondisclosed] secondary neural stem cell (obtained by differentiating neural stem cells expressing i) Pax6, Nestin, Sox1, Sox2, Plzf, ZO1, and BRN2; or ii) Pax6, Sox1, Plzf, and ZO1) as required in claims 1, 53, 55.
Response to argument
Applicants argue amendment overcomes the rejection. Applicants’ argument is not persuasive for reasons set forth above.
C) There is no place in the specification that supports or suggests exposing isolated mammalian neural stem cells to TGFβ1 or a nucleic acid sequence encoding NFIA (promoting NFIA signaling) and obtaining a cell population in which at least 10% of the cells express CD44 or AQP4 as encompassed by claims 53 and 55. Pg 4, lines 18-20, contemplates “glial competent cell markers” as CD44, AQP4, Sox2, or Nestin, but they’re not for reasons set forth above. Pg 4, lines 21-25, says 10%, 20%, 30%, 40%... of the cells obtained express a “glial competent cell marker”, but the examples do not teach obtaining a cell population in which at least 10% of the cells express CD44, AQP4, Sox2, or Nestin as claimed. Fig. 2 and 3 show CD44, AQP4, Sox2, and Nestin expression but does not teach 10% of the cells obtained after “expressing NFIA” express CD44, AQP4, Sox2, or Nestin or are “glial cells”. An adequate written description of a method requires more than a mere statement that it is part of the invention and reference to a potential method for isolating it; what is required is a description of an active step that results in obtaining a population of cells in which at least 10% express CD44or AQP4 and are glial cells or some kind of other neural precursor. Disclosure of no more than that, as in the instant case, is simply a wish to identify a means of doing so.
Response to argument
Applicants argue amendment overcomes the rejection. Applicants’ argument is not persuasive for reasons set forth above.
D) The specification lacks written description for obtaining a cell population containing at least 10% cells that express GFAP after stopping the inducing of NFIA (claim 1), removing the inducer of endogenous NFIA (claim 53), or removing exogenous NFIA (claim 55) . Fig. 5 shows GFAP expression after inducing exogenous NFIA expression under the control of an inducible promoter followed by decreasing NFIA expression and culture in LIF (pg 60, lines 5-11). There is nothing in the specification or art at the time of filing that indicates the cell type obtained after the 2nd step of claim 1, 53, 55 is a population of cells in which at least 10% of are “glial competent cells that express GFAP”. Accordingly, the final product in the 2nd step of claims 1, 53, 55 lacks written description.
Response to arguments
Applicants point to Fig. 5. Applicants’ argument is not persuasive for reasons set forth above.
E) The specification lacks written description for exposing any pluripotent cells to an inhibitor of SMAD to obtain neural stem cells expressing i) Pax6, Nestin, Sox1, Sox2, Plzf, ZO1, and BRN2; or ii) Pax6, Sox1, Plzf, and ZO1 as required in claim 2 other than using SB431542 (a TGFβ/Activin-Nodal inhibitor) and LDN193189 (a BMP inhibitor). Pg 7, lines 15-20, contemplates SMAD inhibitors comprise TGFβ/Activin-Nodal signaling inhibitors such as SB431542 or BMP inhibitors, e.g. LDN193189. Pg 22, describes examples of SMAD inhibitors including Noggin; chordin; Cerberus; follistatin; bambi; antibodies that block activins, nodal, TGFβ or BMPs; LDN193189, or SB431542.
Pg 59, lines 1-5, is limited to culturing pluripotent cells in the presence of LDN193189 and SB431542 such that neural stem cells expressing i) Pax6, Nestin, Sox1, Sox2, Plzf, ZO1, and BRN2; or ii) Pax6, Sox1, Plzf, and ZO1 are obtained.
The specification does not correlate using a combination of a TGFβ/Activin-Nodal inhibitor (SB431542) and a BMP inhibitor (LDN193189) to using a TGFβ/Activin-Nodal inhibitor or BMP inhibitor alone as broadly encompassed by claim 2. The specification does not correlate using the combination of SB431542 and LDN193189 to using Noggin; chordin; Cerberus; follistatin; bambi; or antibodies that block activins, nodal, TGFβ or BMPs as broadly encompassed by claim 2.
While methods of differentiating pluripotent cells into neural stem cells were well-known in the art, the concept of exposing pluripotent cells to any SMAD inhibitor as claimed was not. Accordingly, the concept lacks written description other than culturing mammalian pluripotent cells in the presence of SB431542 and LDN193189 such that neural stem cells are obtained.
Response to arguments
Applicants point to paragraph 124 of the instant application which identifies various compounds that inhibit SMAD signaling in addition to LDN193189 and SB431542, e.g. noggin, chordin, Cerberus…. Applicants’ argument is not persuasive. The specification does not have paragraph number. When referencing the specification, please use page and paragraph number (and line number as necessary) of the original specification. Do not simply use paragraph numbers or refer to the US Patent Application Publication. Pg 22, lines 18-27, discuss SMAD inhibitors, but is not persuasive for reasons set forth above.
F) The specification lacks written description for claim 3 other than culturing mammalian pluripotent cells in the presence of SB431542 or LDN193189 such that neural stems cells are obtained. The specification does not contemplate inducing NFIA at least 8 days after exposing the pluripotent cells to an inhibitor of SMAD as required in claim 3. Accordingly, claim 3 lacks written description other than culturing mammalian pluripotent cells in the presence of SB431542 or LDN193189 such that neural stems cells are obtained.
Response to arguments
Applicants argument is the same as the rejection above which is not persuasive because the specification does not contemplate culturing mammalian pluripotent cells in the presence of SB431542 or LDN193189 for 8 days such that neural stems cells are obtained.
G) Claims 33-35 remain rejected for reasons set forth under claim 1 regarding “SMAD signaling”.
New rejections
H) The specification lacks written description for “glial competent neural stem cell that express CD44 or AQP4” as required in step A) of claims 1, 53, 55 or “glial competent cell that express GFAP” as required in step B) of claims 1, 53, 55. It is unclear how a “glial competent neural stem cell that expresses CD44 or AQP4” differs from any glial neural stem cell that expresses CD44 or AQP4. It is unclear how a “glial competent neural stem cell that expresses GFAP” differs from any “glial neural stem cell that expresses GFAP”. The specification and the art at the time of filing do not teach when glial stem cells or glial cells are “competent”. Accordingly, the concept lacks written description.
The concept of “glial competent cells” that are cortical astrocytes or spinal cord astrocytes” in claim 32 lacks written description other than glial cells that are astrocytes. While glial cells may be astrocytes and astrocytes may be located in the cortex or spinal cord, the location of the astrocytes in the cortex or spinal cord does not further limit the astrocytes. The specification and the art at the time of filing do not teach any structures that distinguish cortical astrocytes or spinal cord astrocytes from any astrocyte. Therefore, it is unclear how “cortical astrocytes or spinal cord astrocytes” further limits the glial cells other than being astrocytes.
35 USC § 102
The rejection of claims 1-3, 29, 30, 32-35, 40, 42, 43 under 35 U.S.C. 102a1 as being anticipated by Anderson (6001654) was withdrawn. Anderson differentiated NSCs in culture in the presence of TGFβ1 for 5-10 days until glial cells expressing GFAP were obtained (col. 24, lines 52-67; claim 7). The conditions of Anderson inherently MUST promote NF1A signaling for 5 days as required in claim 1 because Anderson used TGFβ1 (claim 7) and taught glial cells are obtained after 5-10 days. Anderson did not teach the NSCs comprised a nucleic acid encoding an exogenous NFIA as required in step a) of claim 1 or culturing the cells in LIF as required in step b) of claim 1. Anderson did not teach culturing NSCs in the presence of TGFβ1 followed by culture the cells obtained in the presence of LIF and the absence of TGFβ1 or any other NFIA activator as required in step b) of claim 53. Anderson did not culturing the NSCs in the presence of exogenous NFIA as required in step a) of claim 55 or culturing the cells in the presence of LIF and the absence of exogenous NFIA as required in step b) of claim 55.
35 USC § 103
The rejection of claims 53-55 under 35 U.S.C. 103 as being unpatentable over Robinson (Stem Cells and Development, 2011, Vol. 20, No. 2, pg 289-300) was withdrawn. Robinson differentiated NSCs into glial cells using mesenchymal stem cells (MSCs) that secrete exogenous TGFβ1 after 4 days in culture (pg 294, col. 1, 2nd full paragraph). The MSCs used by Robinson inherently must also express LIF as required in step b) of claims 53 and 55 because a cell population in which 23.6% of the cells expressed GFAP was obtained. Robinson did not teach culturing NSCs in the presence of TGFβ1 followed by culture the cells obtained in the presence of LIF and the absence of TGFβ1 or any other NFIA activator as required in step b) of claim 53. Robinson did not culturing the NSCs in the presence of exogenous NFIA as required in step a) of claim 55 or culturing the cells in the presence of LIF and the absence of exogenous NFIA as required in step b) of claim 55.
Conclusion
The prior art made of record and not relied upon is considered pertinent to applicant's disclosure:
Salimi (Mol Biol Rep, 2014, Vol. 41, pg 1713-1721) taught a protocol for differentiating pluripotent cells into cells expressing Nestin and other neural stem cell-specific markers (pg 1716, col. 1; pg 1718, end of column 1; paragraph bridging pg 1718-1719).
No claim is 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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Michael C. Wilson
/MICHAEL C WILSON/
Primary Examiner, Art Unit 1638
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