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 1-20 are pending.
Election/Restrictions
Applicant’s election without traverse of Group VIII, claims 9-11, in the reply filed on 11-3-25 is acknowledged.
Claims 1-8, 12-20 have been withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim.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 11-3-25.
Claims 9-11 are under consideration.
Claim Rejections - 35 USC § 112
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Written Description
Claims 9-11 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for 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.
The specification lacks written description for any “bladder organoid” containing 1st a layer of cells that express UPK1B or UPK2 but not P63 and a 2nd layer of cells that express P63 and P63 and UPK1B or UPK2 “localized in an outward direction relative to the first layer” as required in claim 9.
Claim 9 is drawn to a “bladder organoid comprising a first cell layer comprising cells that express UPK1B and/or UPK2 and do not substantially express P63; and a second cell layer localized in an outward direction relative to the first cell layer, the second cell layer comprising cells that co-express P63 and UPK1B and/or UPK2;wherein the bladder organoid may further comprise a third cell layer localized in an outward direction relative to the second cell layer, the third cell layer comprising cells that co- express P63 and KRT5”.
The claim encompasses an artificial bladder or two layer of cells without any bladder function. The cells may be from one of any species or a mixture of species. The “organoid” may have any function, including one or more functions of a bladder, or it may have no function. The “organoid” may have “hindgut” tissue or not. The “hindgut” may be “ventralized” or not. The “organoid” may have bladder epithelium or not.
The 1st and 2nd layer of cells may be co-cultured or both be obtained by differentiating pluripotent cells or definitive endoderm.
The 3rd layer is optional.
The structures and functions that define when the 2nd layer is “localized in an outward direction relative to the first cell layer” or when the 3rd optional layer is “localized in an outward direction relative to the second cell layer” cannot be determined.
Vasyutin (Anticancer Res., 2019, Vol. 39, pg 1105-1118) taught “Bladder Organoids and Spheroids: Potential Tools for Normal and diseased Tissue Modeling”.
Ofuji (Program/Proceedings of the Mol. Biol. Soc. of Japan, 2018, Vol. 41, 1P-0434) taught differentiating human iPS cells into definitive endoderm (DE) using activin A and a Wnt agonist. The DE was differentiated into hindgut-like spheroids using FGF-4 and a Wnt agonist. Expression of genes such as P63 was increased when the hindgut-like spheroids were cultured.
Example 1 teaches differentiating human iPS cells into DE using activin A and Wnt agonist CHIR99021 (pg 67, para 176) followed by differentiation into hindgut using CHIR99021 and FGF4 until floating spheres were obtained (para 177). The specification does not teach the floating spheres were bladder organoids because applicants do not teach the spheres shared any structure or function with a bladder or with any specific bladder tissue. The specification does not teach the spheres have a lumen like a bladder. The specification does not teach the spheres have multiple tissue layers found in a bladder. The specification does not teach the spheres express markers of any specific bladder tissue.
The floating spheres were cultured in Matrigel using a membrane until “slightly rounded tubular cellular structures (hindgut organoids)” were obtained (pg 179). The specification does not teach the rounded tubular cellular structures were bladder organoids because applicants do not teach they shared any structure or function with a bladder or with any specific bladder tissue. The specification does not teach the rounded tubular cellular structures have a lumen like a bladder. The specification does not teach the rounded tubular cellular structures have multiple tissue layers found in a bladder. The specification does not teach the rounded tubular cellular structures express markers of any specific bladder tissue. In fact, applicants refer to the rounded tubular cellular structures as “hindgut organoids” and not “bladder organoids”; however, this appears to be contrary to Ofuji who referred to “bladder organoids” obtained by simply differentiating human iPS cells into DE using activin A and a Wnt agonist followed by differentiating into hindgut-like spheroids using FGF-4 and a Wnt agonist to obtain P63+ hindgut-like spheroids.
The rounded tubular cellular structures were “ventralized” by culturing them in media containing BMP4, FGF4, and CHIR99021 to become “spherical cellular structures (ventral hindgut organoids)” (pg 68, para 180). The “spherical cellular structures” expressed P63 and weakly expressed dorsal hindgut markers CDX2 and Sox2 (pg 69, Table 1). The specification does not teach the “spherical cellular structures” were bladder organoids because applicants do not teach they shared any structure or function with a bladder or with any specific bladder tissue. The specification does not teach the “spherical cellular structures” have a lumen like a bladder. The specification does not teach the “spherical cellular structures” have multiple tissue layers found in a bladder. The specification does not teach the “spherical cellular structures” express markers of any specific bladder tissue or had two layers having the marker pattern claimed. In fact, applicants refer to the rounded tubular cellular structures as “ventral hindgut organoids” and not “bladder organoids”; however, this appears to be contrary to Ofuji who referred to “bladder organoids” obtained by simply differentiating human iPS cells into DE using activin A and a Wnt agonist followed by differentiating into hindgut-like spheroids using FGF-4 and a Wnt agonist to obtain P63+ hindgut-like spheroids.
The “spherical cellular structures” were cultured in BMP2, EGF, RSPO1 and retinoic acid) such that “intestine-like organoids” expressing SATB2 were obtained (pg 70, para 183). The “intestine-like organoids” expressed “ventral hindgut/cloaca marker P63” as well as bladder epithelial markers UPK2 and KRT5 (pg 71, para 185). The “intestine-like organoids” expressed “mid/hindgut marker CDX2 in part and expressed an epithelial marker ECAD” (para 184). The specification does not teach the “intestine-like organoids” were bladder organoids because applicants do not teach they shared any structure or function with a bladder or with any specific bladder tissue. The specification does not teach the “intestine-like organoids” have a lumen like a bladder. The specification does not teach the “intestine-like organoids” have multiple tissue layers found in a bladder. The specification does not teach the “intestine-like organoids” express markers of any specific bladder tissue or had two layers having the marker pattern claimed. In fact, applicants refer to the rounded tubular cellular structures as “intestine-like organoids” and not “bladder organoids”; however, this appears to be contrary to Ofuji who referred to “bladder organoids” obtained by simply differentiating human iPS cells into DE using activin A and a Wnt agonist followed by differentiating into hindgut-like spheroids using FGF-4 and a Wnt agonist to obtain P63+ hindgut-like spheroids.
The “spherical cellular structures” were alternatively cultured in BMP4, all-trans retinoic acid, and FGF7 such that “bladder-like organoids” were obtained (pg 71, Example 2, para 188). The “bladder-like organoids” expressed ECAD, UPK2, and KRT5 (pg 72, para 190). Fig. 3D shows “basal cell-like cells co-expressing p63 and KRT5 localized along the outermost periphery of the organoid” and “intermediate cell-like cells co-expressing p63and UPK2 [ ] localized in an inward direction relative to an area where the intermediate cell-like cells are localized” which is “similar to that of the developing bladder epithelium in vivo” (pg 72, para 190). However, the meaning of “intermediate cell-like cells co-expressing p63and UPK2 [ ] localized in an inward direction relative to an area where the intermediate cell-like cells are localized” cannot be determined by looking at Fig. 3D:
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The specification does not teach the “bladder-like organoids” have a lumen like a bladder. The specification does not teach the “bladder-like organoids” have all of the tissue layers found in a bladder. The specification does not teach the “bladder-like organoids” had two layers having the marker pattern claimed or the 3rd layer claimed.
Example 3 (pg 73) teaches culturing human iPS cells “the same way as in Example 1, except for seeding the cells at 90,000 cells/cm2 onto a 6-well plate” followed by induction into DE, induction into hindgut, “ventralizing” the hindgut, and transplanting the tissue obtained “under the renal capsule of immunocompromised mice (pg 73-74; para 192-194). “The transplanted ventral hindgut organoid became a cellular structure (bladder organoid) with a pouch-like structure having a lumen after four weeks of transplantation, like the living bladder. The bladder organoid contained cells expressing the bladder epithelial markers UPK2 (Fig. 5a), P63 (Fig. 5b), and KRT5 (Fig. 5c), which formed a similar structure to the layer structure of the matured bladder in vivo (Fig. 5g). Specifically, the bladder organoid had Krt5- basal cell-like cells co-expressing P63 and KRT5 localized at the outermost periphery of the pouch-like cellular structure (Fig. 5c). In addition, a few Intermediate cell-like cells co-expressing UPK2 and P63 existed in an inward direction relative to the region where the KRT5-basal cell-like cells were localized (Fig. 5f). Furthermore, Superficial cell-like cells, expressing UPK2 but neither expressing P63 nor KRT5, were localized in an inward direction to the region where the Intermediate cell-like cells were localized and to the region facing the lumen of the bladder organoid (Fig. 5e and f).” (pg 74-75, para 196). However, the meaning of “intermediate cell-like cells co-expressing p63and UPK2 [that] existed in an inward direction relative to an area where the KRT5-basal cell like cells are localized” cannot be determined by looking at Fig. 5C:
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The specification does not teach the “bladder organoids” have all of the tissue layers found in a bladder. The specification does not teach the “bladder organoids” had two layers having the marker pattern claimed or the 3rd layer in claim 9 or the 4th or 5th layer in claim 10.
Accordingly, the specification lacks written description for any bladder organoid, specifically one without a lumen or having the 3rd, 4th, and 5th layers optional, as broadly encompassed by claim 9, or the 2nd layer being “localized in an outward direction relative to the 1st layer” or a 3rd layer being “localized in an outward direction relative to the 2nd layer”.
Enablement
Claims 9-11 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, because the specification, while being enabling for a bladder organoid, does not reasonably provide enablement for the bladder organoid having the structures in claim 9. The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make/use the invention commensurate in scope with these claims.
The specification does not enable making/using any “bladder organoid” containing 1st a layer of cells that express UPK1B or UPK2 but not P63 and a 2nd layer of cells that express P63 and P63 and UPK1B or UPK2 “localized in an outward direction relative to the first layer” as required in claim 9.
Claim 9 and its scope are discussed above.
The art at the time of filing is summarized above.
Example 1 teaches differentiating human iPS cells into DE using activin A and Wnt agonist CHIR99021 (pg 67, para 176) followed by differentiation into hindgut using CHIR99021 and FGF4 until floating spheres were obtained (para 177). The specification does not teach the floating spheres were bladder organoids because applicants do not teach the spheres shared any structure or function with a bladder or with any specific bladder tissue. The specification does not teach the spheres have a lumen like a bladder. The specification does not teach the spheres have multiple tissue layers found in a bladder. The specification does not teach the spheres express markers of any specific bladder tissue.
The floating spheres were cultured in Matrigel using a membrane until “slightly rounded tubular cellular structures (hindgut organoids)” were obtained (pg 179). The specification does not teach the rounded tubular cellular structures were bladder organoids because applicants do not teach they shared any structure or function with a bladder or with any specific bladder tissue. The specification does not teach the rounded tubular cellular structures have a lumen like a bladder. The specification does not teach the rounded tubular cellular structures have multiple tissue layers found in a bladder. The specification does not teach the rounded tubular cellular structures express markers of any specific bladder tissue. In fact, applicants refer to the rounded tubular cellular structures as “hindgut organoids” and not “bladder organoids”; however, this appears to be contrary to Ofuji who referred to “bladder organoids” obtained by simply differentiating human iPS cells into DE using activin A and a Wnt agonist followed by differentiating into hindgut-like spheroids using FGF-4 and a Wnt agonist to obtain P63+ hindgut-like spheroids.
The rounded tubular cellular structures were “ventralized” by culturing them in media containing BMP4, FGF4, and CHIR99021 to become “spherical cellular structures (ventral hindgut organoids)” (pg 68, para 180). The “spherical cellular structures” expressed P63 and weakly expressed dorsal hindgut markers CDX2 and Sox2 (pg 69, Table 1). The specification does not teach the “spherical cellular structures” were bladder organoids because applicants do not teach they shared any structure or function with a bladder or with any specific bladder tissue. The specification does not teach the “spherical cellular structures” have a lumen like a bladder. The specification does not teach the “spherical cellular structures” have multiple tissue layers found in a bladder. The specification does not teach the “spherical cellular structures” express markers of any specific bladder tissue or had two layers having the marker pattern claimed. In fact, applicants refer to the rounded tubular cellular structures as “ventral hindgut organoids” and not “bladder organoids”; however, this appears to be contrary to Ofuji who referred to “bladder organoids” obtained by simply differentiating human iPS cells into DE using activin A and a Wnt agonist followed by differentiating into hindgut-like spheroids using FGF-4 and a Wnt agonist to obtain P63+ hindgut-like spheroids.
The “spherical cellular structures” were cultured in BMP2, EGF, RSPO1 and retinoic acid) such that “intestine-like organoids” expressing SATB2 were obtained (pg 70, para 183). The “intestine-like organoids” expressed “ventral hindgut/cloaca marker P63” as well as bladder epithelial markers UPK2 and KRT5 (pg 71, para 185). The “intestine-like organoids” expressed “mid/hindgut marker CDX2 in part and expressed an epithelial marker ECAD” (para 184). The specification does not teach the “intestine-like organoids” were bladder organoids because applicants do not teach they shared any structure or function with a bladder or with any specific bladder tissue. The specification does not teach the “intestine-like organoids” have a lumen like a bladder. The specification does not teach the “intestine-like organoids” have multiple tissue layers found in a bladder. The specification does not teach the “intestine-like organoids” express markers of any specific bladder tissue or had two layers having the marker pattern claimed. In fact, applicants refer to the rounded tubular cellular structures as “intestine-like organoids” and not “bladder organoids”; however, this appears to be contrary to Ofuji who referred to “bladder organoids” obtained by simply differentiating human iPS cells into DE using activin A and a Wnt agonist followed by differentiating into hindgut-like spheroids using FGF-4 and a Wnt agonist to obtain P63+ hindgut-like spheroids.
The “spherical cellular structures” were alternatively cultured in BMP4, all-trans retinoic acid, and FGF7 such that “bladder-like organoids” were obtained (pg 71, Example 2, para 188). The “bladder-like organoids” expressed ECAD, UPK2, and KRT5 (pg 72, para 190). Fig. 3D shows “basal cell-like cells co-expressing p63 and KRT5 localized along the outermost periphery of the organoid” and “intermediate cell-like cells co-expressing p63and UPK2 [ ] localized in an inward direction relative to an area where the intermediate cell-like cells are localized” which is “similar to that of the developing bladder epithelium in vivo” (pg 72, para 190). However, the meaning of “intermediate cell-like cells co-expressing p63and UPK2 [ ] localized in an inward direction relative to an area where the intermediate cell-like cells are localized” cannot be determined by looking at Fig. 3D:
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The specification does not teach the “bladder-like organoids” have a lumen like a bladder. The specification does not teach the “bladder-like organoids” have all of the tissue layers found in a bladder. The specification does not teach the “bladder-like organoids” had two layers having the marker pattern claimed or the 3rd layer claimed.
Example 3 (pg 73) teaches culturing human iPS cells “the same way as in Example 1, except for seeding the cells at 90,000 cells/cm2 onto a 6-well plate” followed by induction into DE, induction into hindgut, “ventralizing” the hindgut, and transplanting the tissue obtained “under the renal capsule of immunocompromised mice (pg 73-74; para 192-194). “The transplanted ventral hindgut organoid became a cellular structure (bladder organoid) with a pouch-like structure having a lumen after four weeks of transplantation, like the living bladder. The bladder organoid contained cells expressing the bladder epithelial markers UPK2 (Fig. 5a), P63 (Fig. 5b), and KRT5 (Fig. 5c), which formed a similar structure to the layer structure of the matured bladder in vivo (Fig. 5g). Specifically, the bladder organoid had Krt5- basal cell-like cells co-expressing P63 and KRT5 localized at the outermost periphery of the pouch-like cellular structure (Fig. 5c). In addition, a few Intermediate cell-like cells co-expressing UPK2 and P63 existed in an inward direction relative to the region where the KRT5-basal cell-like cells were localized (Fig. 5f). Furthermore, Superficial cell-like cells, expressing UPK2 but neither expressing P63 nor KRT5, were localized in an inward direction to the region where the Intermediate cell-like cells were localized and to the region facing the lumen of the bladder organoid (Fig. 5e and f).” (pg 74-75, para 196). However, the meaning of “intermediate cell-like cells co-expressing p63and UPK2 [that] existed in an inward direction relative to an area where the KRT5-basal cell like cells are localized” cannot be determined by looking at Fig. 5C:
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The specification does not teach the “bladder organoids” have all of the tissue layers found in a bladder. The specification does not teach the “bladder organoids” had two layers having the marker pattern claimed or the 3rd layer in claim 9 or the 4th or 5th layer in claim 10.
Given the lack of guidance in the specification taken with the art at the time of filing, it would have required those of skill undue experimentation to determine how to make/use any bladder organoid, specifically one without a lumen or having the 3rd, 4th, and 5th layers optional, as broadly encompassed by claim 9, or the 2nd layer being “localized in an outward direction relative to the 1st layer” or a 3rd layer being “localized in an outward direction relative to the 2nd layer”.
Indefiniteness
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 9-11 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.
The concept of a 2nd layer of cells “localized in an inward direction relative to the first cell layer” in claim 9 makes the claim indefinite because the metes and bounds cannot be determined. Example 2 taught making “bladder-like organoids” that expressed ECAD, UPK2, and KRT5 (pg 72, para 190). Fig. 3D shows “basal cell-like cells co-expressing p63 and KRT5 localized along the outermost periphery of the organoid” and “intermediate cell-like cells co-expressing p63and UPK2 [ ] localized in an inward direction relative to an area where the intermediate cell-like cells are localized” which is “similar to that of the developing bladder epithelium in vivo” (pg 72, para 190). However, the meaning of “intermediate cell-like cells co-expressing p63and UPK2 [ ] localized in an inward direction relative to an area where the intermediate cell-like cells are localized” cannot be determined by looking at Fig. 3D:
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The metes and bounds of the 1st and 2nd layers being “localized” cannot be determined. In fact, it appears the 2nd layer expressing p63 and UPK1B or UPK2 is on the inside of the sphere.
Pg 74-75, para 196, is cited above and says “intermediate cell-like cells co-expressing p63and UPK2 [that] existed in an inward direction relative to an area where the KRT5-basal cell like cells are localized” cannot be determined by looking at Fig. 5C:
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However, the metes and bounds of the 1st and 2nd layers being “localized” cannot be determined as required in claim 9.
Similarly, the metes and bounds of a 3rd layer being “localized in an outward direction relative to the second layer” in claim 9, a 4th or 5th layer being “localized outward relative to the [3rd or 4th] layer” in claim 11 cannot be determined for reasons cited above.
Vasyutin (Anticancer Res., 2019, Vol. 39, pg 1105-1118) taught “Bladder Organoids and Spheroids: Potential Tools for Normal and diseased Tissue Modeling”, so any of the tissues in Examples 1-3 can be used as a tool for tissue modeling.
Ofuji (Program/Proceedings of the Mol. Biol. Soc. of Japan, 2018, Vol. 41, 1P-0434) taught differentiating human iPS cells into definitive endoderm (DE) using activin A and a Wnt agonist. The DE was differentiated into hindgut-like spheroids using FGF-4 and a Wnt agonist. Expression of genes such as P63 was increased when the hindgut-like spheroids were cultured. However, the expression pattern claimed does not appear to apply to any of the tissues obtained by applicants in Examples 1 or 2 which all start with the method of Ofuji. It appears that the organoid in claim 9 is attempting to capture the tissue obtained after being implanted into a mouse in Example 3 and the expression pattern in claim 9 does not occur until then. The specification and the art at the time of filing do not teach any other way to obtain the marker pattern claimed; therefore, the art at the time of filing did not reasonably teach or suggest the bladder organoid of claim 9.
Conclusion
No claim is allowed.
Inquiry concerning this communication or earlier communications from the examiner should be directed to Michael C. Wilson who can normally be reached at the office on Monday through Friday from 9:30 am to 6:00 pm at 571-272-0738.
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Michael C. Wilson
/MICHAEL C WILSON/
Primary Examiner, Art Unit 1638