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 .
Priority
The instant application is a national stage entry under 35 U.S.C. § 371 of PCT/GB2022/052717 (filed 10/25/2022). Acknowledgement is made of Applicants’ claim for priority to foreign application GB2115337.4 (filed 10/25/2021).
Election/Restrictions
Applicant’s election without traverse of Group II, a method of making a construct for mimicking an in vivo environment, in the reply filed on 06/29/2026 is acknowledged.
Claims 1-2, 4, 7-10, 12, 14, 18-20, 22-28, and 30 are pending in the instant application; claims 1-2, 4, 7-10, 12, 14, 18, and 30 are withdrawn as directed to inventions non-elected.
Claims 19-20 and 22-28 read on the elected invention and are examined on the merits herein.
Claim Objections
Claims 19-20 and 26-27 are objected to because of the following informalities:
Regarding claims 19-20: These claims are directed to a method of making a construct for mimicking an in vivo environment. Claim 19 recites the construct as comprising components recited in (a), (b), (c), and (f); claim 2 further recites components (d), (e), (g), and (h). The incorrect order of the letters designating the various components creates confusion. Further, claim 27 recites “…optionally one or more of cells (e) to (h)…”, which renders the claim indefinite as a direct result of the incorrect order of the letters (discussed below).
Regarding claim 26: This claim does not end with a period. Additionally, line 1 recites “…method of claim 19 further comprising…”; this should read “…method of claim 19, further comprising…” for consistency with the formatting of the other pending claims.
Regarding claim 27: Line 3 contains a typographical error, as there is an extra parenthesis after “(h)”.
Appropriate correction is required.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 19-20 and 22-28 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.
Regarding claim 19: The phrase “such as” recited in line 1 of claim 19 renders the claim indefinite because it is unclear whether the limitations following the phrase are part of the claimed invention. See MPEP § 2173.05(d).
Regarding claim 22: The phrase “(e.g., bioprinting)” recited in line 2 of claim 22 renders the claim indefinite because the “e.g.” portion of this phrase is exemplary claim language; it is unclear whether the limitation following “e.g.” is part of the claimed invention. See MPEP § 2173.05(d). Additionally, while parentheticals can be used for abbreviations, the use of parentheticals adjacent to terms which recite exemplary embodiments renders the claim indefinite because it is not clear whether the items within the parentheticals are required or optional.
Additionally, claim 22 recites the limitation “the extracellular matrix”. There is insufficient antecedent basis for this limitation in the claim. Claim 22 depends from claim 19, but the optional “extracellular matrix” component is not recited until claim 20.
Regarding claim 23: This claim recites the limitation “The method of claim 19, comprising printing…”. There is insufficient antecedent basis for the “printing” limitation in the claim. For purposes of examination, the printing limitation is interpreted as referring to the manner of depositing the cells, akin to the clearly defined metes and bound of claim 22.
Regarding claim 24: This claim further narrows the scope of the method of claim 19 with the Markush grouping of alternative embodiments recited in (i), (ii), and (iii); these embodiments are directed to “depositing or printing…on to the surface…”. Are the “depositing or printing” steps of the instant claim meant to convey wherein the depositing comprises depositing or printing the cells? For purposes of examination, the depositing or printing limitations are interpreted as referring to depositing the cells, wherein the depositing optionally comprises printing.
Claim 24 further lacks clarity regarding the spheroid limitations. Embodiment (i) is directed to “depositing or printing spheroids”; embodiment (ii) is directed to “depositing or printing cells and then subsequently culturing the cells to form spheroids”; embodiment (iii) is directed to “depositing or printing a mixture of spheroids and single cells”. Do the spheroids and/or cells comprise the cells of (a), (b), (c), and optionally (f)? Is there antecedent basis for the spheroids, or do they comprise an entirely different cell type? As currently written, the metes and bounds of claim 24 are not clearly or precisely defined, rendering the claim indefinite. For purposes of examination, the spheroids and/or cells are interpreted as comprising the cells of (a), (b), (c), and optionally (f).
Regarding claim 27: This claim recites the limitation “The method of claim 19, comprising printing…”. There is insufficient antecedent basis for the “printing” limitation in the claim. For purposes of examination, the printing limitation is interpreted as referring to the manner of depositing the cells, akin to the clearly defined metes and bound of claim 22.
Further, there is insufficient antecedent basis for the “…optionally one or more of cells (e) to (h)…” and “…the extracellular matrix (d)” limitations in the claim, as claim 27 depends from claim 19, and components (d), (e), (g), and (h) are not recited until claim 20.
Additionally, claim 27 recites, “…wherein the bioink formulation is made up of the cells (a) to (c) and optionally one or more of cells (e) to (h)) that have been pre-mixed with and/or are suspended in the extracellular matrix (d).” The extra parenthesis is discussed in the Claim Objections section above. However, this results in a lack of clarity, as it is unclear whether the parenthesis in question is intended to be a comma.
The scope of comprising printing droplets of a bioink formulation on to the surface, wherein the bioink formulation is made up of the cells (a) to (c) and optionally one or more of cells (e) to (h), that have been pre-mixed with and/or are suspended in the extracellular matrix (d) is much narrower than the scope of comprising printing droplets of a bioink formulation on to the surface, wherein the bioink formulation is made up of the cells (a) to (c) and optionally one or more of cells (e) to (h) that have been pre-mixed with and/or are suspended in the extracellular matrix (d). The scope of the latter encompasses printing bioink droplets comprising cells (a) to (c), with an optional embodiment of the bioink further comprising cells (e) to (h) pre-mixed with and/or suspended in extracellular matrix; the scope of the former encompasses printing bioink droplets comprising cells (a) to (c) pre-mixed with and/or suspended in extracellular matrix, with an optional embodiment of including cells (e) to (h) in the bioink.
Thus, the metes and bounds of claim 27 are not clearly or precisely defined, rendering the claim indefinite. For purposes of examination, the broadest reasonable interpretation of the limitations recited in claim 27 is printing bioink droplets comprising cells (a) to (c).
Regarding claim 28: There is insufficient antecedent basis for “[the] spheroids” and “the extracellular matrix” limitations recited in this claim; neither the spheroids of claim 24 nor the extracellular matrix of claim 20 are in the chain of dependency for claim 28.
Claims 20 and 25-26 depend from one or more of the above claims, inherit the deficiencies, and are likewise rejected as indefinite.
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 19-20, 24, and 26 are rejected under 35 U.S.C. 102(a)(1) and (a)(2) as being anticipated by Ajji, et al. (WO 2021/087613).
Ajji, et al. teaches a three-dimensional cell culture system (Abstract).
Regarding claims 19-20: Ajji, et al. teaches a method for preparing a three-dimensional (3D) cell culture system, the method comprising providing a functionalized solid porous matrix support made of a biocompatible material; seeding a first type of cells on the support to attach the cell type thereto; and subsequently contacting the support with a biocompatible hydrogel comprising a second type of cell, thereby obtaining the 3D cell culture system (par. 00151). The first type of cell may be any type of cell suitable to mimic a tissue or organ of interest; in an embodiment, the first type of cell comprises a combination of endothelial cells, epithelial cells, and immune cells (par. 00134). The second type of cells comprises at least one type of cancer cell and at least one type of non-cancer cells which are found in a tumor but not the malignant cancer cells per se; e.g., cancer-associated fibroblasts (par. 00144). In an embodiment, the second type of cells further comprises tumor stem-like cells (par. 0046); additionally, the system may further comprise extracellular matrix (pars. 00136, 00145).
For clarity of record, the instant specification states a supportive cell functions to provide structural support to the construct, e.g., epithelial cells (pg. 4; lines 28-30); thus, the epithelial cell reads on the supportive cell of claim 20.
Therefore, the 3D cell culture system of Ajji, et al. anticipates:
the method of making a construct for mimicking an in vivo environment, such as an in vivo cancer microenvironment, wherein the method comprises depositing on a surface: (a) a cancer cell; (b) a cancer stem cell; and (c) a cancer associated fibroblast limitations recited in claim 19; and
the further comprising depositing one or more of the following: (d) an extracellular matrix; (e) a supportive cell; (g) an immune cell; and (h) an endothelial cell limitations recited in claim 20.
Regarding claim 24: Following the above discussion, Ajji, et al. teaches an embodiment wherein the cells within the 3D cell culture system form spheroids (par. 00149); this anticipates the depositing cells on to the surface and then subsequently culturing the cells to form spheroids within the deposited cells limitation recited in part (ii) of claim 24.
Regarding claim 26: Following the above discussion, Ajji, et al. teaches the 3D cell culture system is useful in therapeutic compound screening, such as the evaluation of a compound’s ability to inhibit the migration of tumor or cancer cells; i.e., metastasis (par. 00159). Further disclosed is an embodiment wherein the 3D cell culture system is cultured under conditions to allow the second type of cells (i.e., the cancer cells) to migrate to the surface and/or into the solid porous support (par. 00153); i.e., mimicking metastasis. Therefore, this anticipates the further comprising culturing the construct for a period of time such that the construct mimics an in vivo cancer microenvironment limitation recited in claim 26.
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.
Claims 19-20, 22-24, and 26-28 are rejected under 35 U.S.C. 103 as being unpatentable over Sears, et al. (US 2016/0040132) in view of Li, et al. (Cancer Res. 2007), as evidenced by Allam, et al. (Pancreatology. 2017).
Sears, et al. teaches three-dimensional bioprinted pancreatic tumor tissue structures (Abstract).
Li, et al. teaches pancreatic cancer stem cells (Abstract).
Regarding claims 19-20, 22, 26: Sears, et al. teaches a method of fabricating a three-dimensional, engineered, pancreatic tumor model for recapitulating the pancreatic tumor microenvironment, the method comprising: preparing a stromal bio-ink comprising pancreatic stellate cells and endothelial cells; preparing a tumor bio-ink comprising pancreatic cancer cells; bioprinting the stromal bio-ink and the tumor bio-ink such that the tumor bio-ink is encased in the stromal bio-ink and in contact with the stromal bio-ink on all sides; and maturing the deposited bio-ink in a cell culture media to allow the cells to cohere to form a three-dimensional, engineered, biological tumor model (pars. 0003, 0008) on a biocompatible surface (pars. 0049, 0050). In an embodiment, the bio-ink comprises immune cells and cancer associated fibroblasts (par. 0008); additionally, the bio-ink may further comprise extracellular matrix (par. 0059).
As set forth above, the instant specification states a supportive cell functions to provide structural support to the construct (pg. 4; lines 28-30); the specification states a specialized cell type provides a specific microenvironment in accordance with the type of cancer microenvironment the structure is intended to mimic or reflect (pg. 4; lines 33-35). As evidenced by Allam, et al. (Pancreatology. 2017), pancreatic stellate cells are stromal cells whose physiological functions include the production of extracellular matrix; in pancreatic cancer, the stellate cells exhibit a myofibroblastic-like morphology wherein this activated form engages in several mechanism which support tumorigenesis and cancer invasion and progression (Abstract). Thus, the pancreatic stellate cells read on both the specialized cell type of claim 19 and the supportive cell of claim 20.
Therefore, the pancreatic tumor model of Sears, et al. reads on:
the method of making a construct for mimicking an in vivo environment, such as an in vivo cancer microenvironment, wherein the method comprises depositing on a surface: (a) a cancer cell; (c) a cancer associated fibroblast; and (f) a specialized cell type to provide a specific microenvironment limitations recited in claim 19;
the further comprising depositing (d) an extracellular matrix; (e) a supportive cell; (g) an immune cell; and (h) an endothelial cell limitation recited in claim 20;
the wherein the step of depositing comprises printing (e.g., bioprinting) the cells, and the extracellular matrix, on to the surface limitations recited in claim 22; and
the further comprising culturing the construct for a period of time such that the construct mimics an in vivo cancer microenvironment limitation recited in claim 26.
Sears, et al. does not teach the bio-ink as comprising a cancer stem cell, as required by the remaining limitation of claim 19.
However, Li, et al. teaches a subpopulation of CD44+CD24+ESA+ pancreatic cancer cells which are highly tumorigenic (Table 2; Fig. 3C-D). Li, et al. discloses these cells were injected into mice and the resultant tumors were analyzed, wherein the cells produced additional CD44+CD24+ESA+ cells as well as phenotypically diverse nontumorigenic cells with the same phenotype complexity as the primary tumor from which the cells were derived (pg. 1034); as the tumors were passaged through four rounds of tumor formation with similar results and no decrease in tumorigenicity, Li, et al. teaches these CD44+CD24+ESA+ cells function as cancer stem cells, capable of self-renewal and generating phenotypically diverse progeny (pg. 1034; pg. 1035, par. 1).
Therefore, it would have been prima facie obvious to a person having ordinary skill in the art to have modified the method of Sears, et al by including the CD44+CD24+ESA+ pancreatic cancer stem cells of Li, et al. in the bio-ink for fabricating the pancreatic tumor model. This conclusion of obviousness is based on the ‘teaching, suggestion, or motivation rationale’. Sears, et al. teaches the pancreatic tumor model of the disclosure is advantageous for creating a model to explore the unique biology of pancreatic adenocarcinoma tumors and identification of therapeutic compounds therefor, a distinct advantage over current experimental testing systems which do not accurately recapitulate the composition of the pancreatic tumor microenvironment (pars. 0003, 0006); the CD44+CD24+ESA+ pancreatic cancer stem cells of Li, et al. are derived from primary human pancreatic adenocarcinomas (pg. 1031; col. 1, par 2). Thus, the skilled artisan would be motivated to include the CD44+CD24+ESA+ pancreatic cancer stem cells in the bio-ink in order to more accurately recapitulate the pancreatic tumor microenvironment. Further, as Sears, et al. already teaches the inclusion of pancreatic cancer cells in the bio-ink, the skilled artisan would have more than a reasonable expectation of success.
This renders obvious the remaining wherein the method comprises depositing on a surface: (b) a cancer stem cell limitation recited in claim 19.
Regarding claim 23: Following the above discussion, Sears, et al. teaches an embodiment wherein a plurality of the 3D, engineered pancreatic tumor models are deposited into wells of a multi-well plate to form an array (pg. 2; par. 0010), and wherein the tumor models of the array comprise multiple elements spatially arranged in a pre-determined pattern (par. 0074). This reads on the printing a plurality of constructs at a series of defined and/or discrete locations on a surface, optionally in a predetermined pattern to provide an array or microarray of constructs limitation recited in claim 23.
Regarding claim 24: Following the above discussion, Sears, et al. teaches an embodiment wherein the engineered pancreatic tumor tissues are bioprinted in the form of a sphere, as well as an alternative embodiment wherein the tissues form spheres after a period of maturation in cell culture conditions (par. 0085). This reads on the depositing or printing spheroids on to the surface limitation recited in part (i) as well as the depositing or printing cells on to the surface and then subsequently culturing the cells to form spheroids within the deposited cells or printed cells limitation recited in part (ii) of the instant claim.
Regarding claim 27: Following the above discussion, Sears, et al. teaches an embodiment wherein the bioprinting method is a continuous bioprinting method wherein the bio-ink is dispensed from a bioprinter via a dispense tip with an inner diameter of 20 µm (pars. 0051, 0053); this reads on the printing droplets of a bioink formulation on to the surface, wherein the bioink formulation is made up of the cells (a) to (c) limitation recited in claim 27.
Regarding claim 28: Following the above discussion, Sears, et al. teaches an embodiment wherein between 30% and 100% of the bio-ink comprises cells (par. 0056); this renders obvious the wherein between about 45% and 75% of the total volume of the bioink formulation is comprised of the suspension of cells and/or spheroids limitations recited in claim 28.
Claim 25 is rejected under 35 U.S.C. 103 as being unpatentable over Sears, et al. (US 2016/0040132) in view of Li, et al. (Cancer Res. 2007), further in view of Leonard and Godin (Methods Mol Biol. 2016), as evidenced by Allam, et al. (Pancreatology. 2017).
The teachings of Sears, et al. and Li, et al. are set forth above.
Leonard and Godin (hereinafter Leonard) teaches magnetic manipulation of tumor cells and cells in the tumor microenvironment (Abstract).
Regarding claim 25: Following the above discussion, Sears, et al. does not teach the magnetic bioprinting limitation required by the instant claim.
However, Leonard teaches an important element for in vitro grown spheroids is the presence of a scaffold which mimics the extracellular matrix, an important element in the tumor stroma; such a scaffold can stunt cell growth and affect cell-cell interactions, and the static concentration of the scaffold can cause a misrepresentation in the growing in vivo environment over time (pg. 240; par. 2). Leonard discloses the use of Nanoshuttles® for magnetizing cells to be included in the spheroid, wherein cells are incubated with the Nanoshuttles® and subsequently 3D bioprinted over a magnet, which aids the attraction of the Nanoshuttle® internalized cells to form a 3D tumor mass (Fig. 1). Fibroblasts produce fibronectin and collagen which can naturally form the fibrotic capsule in 3D; fibronectin concentration can increase along with growth of the spheroids containing fibroblasts, enabling a more realistic prognosis for the response of the tumor in vivo to tested drugs (pg. 240; par. 2). Thus, the magnetic bioprinting method enable the simulation of an in vivo environment without using an artificial scaffold and without the need of external surface for support (pg. 240; par. 3).
Therefore, it would have been prima facie obvious to a person having ordinary skill in the art to have further modified the method of Sears, et al. by using magnetic bioprinting, as taught by Leonard. This conclusion of obviousness is based on the ‘teaching, suggestion, or motivation rationale’. One would be motivated to do so to form a realistic 3D tumor spheroid without misrepresenting the tumor microenvironment with the presence of a scaffold, and without the need of external surface for support, as taught by Leonard (pg. 240); further, as Sears, et al. teaches the inclusion of stromal cells such as fibroblasts in the engineered pancreatic tumor model (par. 0091), the skilled artisan would have more than a reasonable expectation of success in doing so.
This renders obvious the further comprising magnetic bioprinting limitation recited in claim 25.
Conclusion
Any inquiry concerning this communication or earlier communications from the examiner should be directed to GINA PRONZATI whose telephone number is (571)270-5725. The examiner can normally be reached Monday - Friday 9:00a - 5:00p ET.
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/GINA PRONZATI/Examiner, Art Unit 1633
/ALLISON M FOX/Primary Examiner, Art Unit 1633
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