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 .
Applicant’s amendments to the claims and arguments filed on 10-29-2025 have been received and entered. Claims 1, 6-7, 13 have been amended. Claims 4-5, 18-20 have been canceled. Claims 21-25 have been added. Claims 1-3, 6-14, 15-17, 21-25 are pending.
It is also noted that since claims 4-5, 18-20 submitted on 10-29-2025 have been canceled, applicants should submit cancelled claims with just the claim number(s) and claim status (cancelled) without any other text for the claims. As per MPEP 714(II)(C)(c): A claim being canceled must be indicated as "canceled;" the text of the claim must not be presented. Providing an instruction to cancel is optional. Canceled and not entered claims must be listed by only the claim number and status identifier, without presenting the text of the claims. When applicant submits the text of canceled or not-entered claims in the amendment, the Office may accept such an amendment, if the amendment otherwise complies with 37 CFR 1.121, instead of sending out a notice of non-compliant amendment to reduce the processing time.
As per 37 C.F.R. 1.121(c)(4):
(4) When claim text shall not be presented; canceling a claim.
(i) No claim text shall be presented for any claim in the claim listing with the status of "canceled" or "not entered."
(ii) Cancellation of a claim shall be effected by an instruction to cancel a particular claim number. Identifying the status of a claim in the claim listing as "canceled" will constitute an instruction to cancel the claim.
Election/Restrictions
Applicant’s election without traverse of Group I (claims 1-14) in the reply filed on 02-05-2025 is acknowledged.
Claims 15 – 20 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected subject matter, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 02-05-2025.
Claims 1-3, 6-14, 21-25 are under consideration.
Priority
This application is a 371 of PCT/EP2020/084092 filed on 12/01/2020 which claims priority from foreign application EP 19213642.2 filed on 12/04/2019 and EP 20178270.3 filed on 06/04/2020. Receipt is acknowledged of certified copies of papers required by 37 CFR 1.55.
Information Disclosure Statement
The information disclosure statements (IDSs) submitted on 10-29-2025 is in compliance with the provisions of 37 CPR 1.97. Accordingly, the information disclosure statement has been considered by the examiner.
Withdrawn-Claim Rejections - 35 USC § 102
Claims 1-3, 12-13 are rejected under 35 U.S.C. 102 (a)(1) as being anticipated by Fisher et al (Adv. Funct. Mater. 2015, 25, 7163–7172, DOI: 10.1002/adfm.201502778). In view of Applicants' amendment of base claim 1, the previous rejections of claims are hereby withdrawn. Applicants' arguments with respect to the withdrawn rejections are thereby rendered moot. The claims are however subject to new rejections over the prior art of record, as set forth below.
Claim Objections
Claim 25 is objected to because of the following informalities:
The term “Pawherein” in line 3 of claim 25 should read “Pa wherein”
Appropriate correction is required.
Maintained in modified form and new- Claim Rejections - 35 USC § 103- necessitated by amendments
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.
Claims 1-3, 6-14 are rejected under 35 U.S.C. 103 as being unpatentable over Fisher et al (Adv. Funct. Mater. 2015, 25, 7163–7172, DOI: 10.1002/adfm.201502778) in view of Lutolf et al (Pub. No.: US 2018/0258403 A1, Pub. Date : Sep.13, 2018) as evidenced by Salgia (Semin Oncol. 2009 April; 36(2 Suppl 1): S52–S58. doi:10.1053/j.seminoncol.2009.02.008.).
Claim interpretation:
The specification of the claimed invention teaches that an array is a set of several discrete volumes that can be arranged in a certain manner, for example in rows and/or columns. For example, a typically used well plate (e.g. a 48-well plate) provides 48 discrete volumes that are arranged in 8 columns and 6 rows, wherein in this example each column consist of 6 discrete volumes. Each such column in this example is considered as an array, according to the present invention (Page 22, lines 14-20). Thus, array with discrete volumes is interpreted as multi-well plate.
According to the present invention, the term "performing an operation with the cells grown" includes the addition of one or more drugs to said discrete volumes of said hydrogel matrix array. The term "performing an operation with the cells grown" includes also operations where the cells themselves or by-products from the cells are analyzed , as well as operations where products of these cells are analyzed or cell-derived products are isolated for further use (Page 28, lines 5-21). Thus, the term "performing an operation with the cells grown" is interpreted as analyzing cells themselves or by-products from the cells or the addition of one or more chemical compounds/drugs.
According the specification of the claimed invention, the term "pre-selected" means that the extracellular matrix conditions, i.e. at least one of said hydrogel precursor molecules, said optional crosslinking agent, said optional bioactive agent, and preferably said culture media, preferably at least two of them and most preferably all of them, are selected for the tissue type to be tested such that a specific combination of hydrogel features has been pre-selected. Hydrogel features are features that define the structure and/or function of a hydrogel (Page 28, lines 23-31). Thus, the phrase “a specific combination of hydrogel features has been pre-selected” in claim 1 is interpreted as extracellular matrix conditions has been pre-selected.
The specification of the claimed invention teaches that for some organs, patient-derived cancer cells that form tumor organoids tend to grow ex vivo more slowly than their healthy (wild-type) counterparts or associated stromal cells (page 97, lines 2-4). The tissue type is cancer cells that grow ex vivo more slowly than their healthy/normal counterparts (e.g. epithelial and/or stromal cells), preferably prostate cancer cells (page 17, lines 19-21). Thus, prostate cancer cells are interpreted as cancer cells that grow ex vivo more slowly than their healthy/normal counterparts.
Regarding to claims 1-3, Fisher et al teach cell screening to study cancer cell invasion, and Hyaluronic acid (HA)-based hydrogels crosslinked with matrix-metalloproteinase (MMP)-cleavable peptides are developed to study MDA-MB-231 breast cancer cell invasion (Abstract).
Fisher et al teach 2×104 cancer cells per well were seeded on the hydrogel surface in 8 well chamber slides (Page 7171, right column, 3rd para.). The hyaluronic acid (HA)-based hydrogels crosslinked with matrix-metalloproteinase (MMP)-cleavable peptides are developed to study MDA-MB-231 breast cancer cell invasion. Hydrogels are synthesized by reacting furan-modified HA with bismaleimide peptide crosslinkers in a Diels–Alder click reaction. Increased crosslink density correlates with decreased MDAMB-231 cell invasion whereas incorporation of MMP-cleavable sequences within the peptide crosslinker enhances invasion (Abstract) (For claim 1, step a, and claim 3).
Fisher et al teach that the cells were cultured on the hydrogels for 4d before being fixed with 4% paraformaldehyde and stained with DAPI for visualization of cell nuclei (Page 7171, right column, 3rd para.) (For claim 1, step b).
Fisher et al teach determining the effects of tuning the hydrogels’ crosslink density, proteolytic degradation of the crosslinker, and ligand density on the invasion of MDA-MB-231 cells by hydrogel modifications: 1) To vary crosslink density, 1.25% w/v HA/MMPx hydrogels were formed from HA with varying furan substitutions. 2) To determine the influence of proteolytic degradation of the peptide crosslinker on invasion, medium crosslink density, 1.00% w/v HA hydrogels were formed with either MMPx or GAGx. 3) To confirm the influence of MMP-dependent invasion, MDAMB-231 cells were cultured on 1.25% w/v HA/MMPx. Hydrogels were preincubated in media, and media were changed daily throughout the experiment. 4) To vary ligand density, prior to neutralizing HA/MMPx, pendant mal-GRGDS was dissolved in MES buffer and soaked into medium crosslink density, HA/MMPx hydrogels overnight (Page 7171, right column, 4th para.) (For claim 1, step c, and the claimed: (i) a specific combination of hydrogel features has been pre-selected for the said one tissue type to be tested and claim 2).
Although Fisher et al teach cell screening to study cancer cell invasion, Fisher et al do not teach: drug screening and using cells from a biopsy and one or more drugs are added (claim 1); lung cancer cells (claim 6), pancreatic ductal adenocarcinoma and PEG hydrogel having a stiffness in the range of 50 to 3000 Pa (claim 8); R-spondin and Wnt 3a (claim 9); colorectal cancer and PEG hydrogel having at least an initial stiffness in the range of 50 to 2000 Pa (claim 10); and cancer cells that grow ex vivo more slowly than their normal counterparts (claim 14). Lutolf et al who teach three dimensional hydrogels for culturing organoids (title) cure the deficiency.
Lutolf et al provides a method for screening of libraries of pharmacologic compounds, biomolecules or evaluating cell-based therapies for efficacy in inducing tumor cell death or growth arrest, the method comprising i) encapsulating tumor cells or organoids in the three-dimensional hydrogel of the invention and culturing the cells or organoids under suitable conditions in the presence of the pharmacologic compounds, biomolecules or cells to be tested, and ii) monitoring cell death and/ or growth arrest ([0034], page 3). Lutolf et al teach that cells are isolated from a biopsy sample, and the cells are cultured in the presence or absence of the pharmacologic compounds or biomolecules to be tested (Page 21, claim 32).
Therefore, it would have been prima facie obvious for a person of ordinary skill in the art before the effective filing date of the rejected claims to combine the teachings of prior art to modify the method of Fisher et al by using a method for screening of libraries of pharmacologic compounds with three dimensional hydrogels for culturing organoids as taught by Lutolf et al as instantly claimed, with a reasonable expectation of success. Said modification amounting to combining prior art elements according to known methods to yield predictable results. One of ordinary skill in the art would have been motivated to do so because Lutolf et al provide several advantages such as AG73-conjugated PEG matrices significantly enhance the growth of organoids ([0038], page 3); Human patient-derived colorectal cancer organoids grow efficiently in soft and stiff matrices, with or without RGD ([0046], page 4); Laminin alone effectively supported colony formation in soft matrices ([0167], page 13). One of ordinary skill in the art would have had a reasonable expectation of success in doing so because Lutolf et al were successfully generated functionalized hydrogels for various cell types for screening of libraries of pharmacologic compounds, with detailed instructions, working examples and data.
Regarding to claim 6, Lutolf et al teach the tumor-derived single cells are derived from lung tumors ([0104], page 8). It is noted that c-Met is inherently overexpressed in lung cancer cell as evidenced by Salgia (Semin Oncol. 2009 April; 36(2 Suppl 1): S52–S58.). Also, Lutolf et al teach FIG.7A shows colony formation efficiency of ISCs embedded in degradable (DG) or non-degradable (N-DG) PEG gels of varying stiffness ([0043], page 4). FIG.10B: Human patient-derived colorectal cancer organoids grow efficiently in soft and stiff matrices, with or without RGD ([0046], page 4). Thus, a person of ordinary skill in the art would be able to use non-degradable (N-DG) PEG gels with or without RGD for cells derived from lung tumors.
Regarding to claim 7, Lutolf et al teach cell culture with the use of R-spondin ([0189], Page 16).
Regarding to claim 8, Lutolf et al teach FIG. 9A: Mouse pancreatic ducts embedded in PEG -RGD ([0045], page 4) and the tumor-derived single cells can be derived from pancreatic tumors ([0104], page 8). Also, FIG.7A shows colony formation efficiency of ISCs embedded in degradable (DG) or non-degradable (N-DG) PEG gels of varying stiffness ([0043], page 4), and the three-dimensional hydrogels of the invention have an initial shear modulus (stiffness) of 0.5-5 kPa, preferably 0.5-2.5 kPa and a final shear modulus (stiffness) of 50-200 Pa ([0086], page 7). Additionally, FIG.9 shows culture and expansion of various mouse and human organoids in PEG-RGD ([0045], page 4). Thus, a person of ordinary skill in the art would be able to use non-degradable (N-DG) PEG gels of varying stiffness of 0.5-5 kPa or 50-200 Pa with a RGD motif.
Regarding to claims 9 and 11, Lutolf et al teach cell culture with the use of R-spondin ([0189], Page 16) and FIG. 3K and L show ISC expansion in Wnt3a -containing medium ([0039], page 3).
Regarding to claim 10, Lutolf et al teach FIG. 10B: Human patient-derived colorectal cancer organoids grow efficiently in soft and stiff matrices, with or without RGD ([0046], Page 4). The hydrogel comprises a crosslinked hydrophilic polymer and a bioactive molecule, wherein the functional molecule is laminin-111 or a functional variant thereof, and wherein the hydrogel has a shear modulus between 0.05 -0.5 kPa ([0024], page 2). Thus, a person of ordinary skill in the art would be able to use patient-derived colorectal cancer organoids in matrices, with or without RGD, with laminin-111 that have a shear modulus between 0.05 -0.5 kPa.
Regarding to claim 12, Fisher et al teach Hyaluronic acid (HA)-based hydrogels crosslinked with matrix-metalloproteinase (MMP)-cleavable peptides are developed to study MDA-MB-231 breast cancer cell invasion (Abstract). Also, HA–furan hydrogels crosslinked with bismaleimide PEG are degradable via hyaluronidase and support the attachment and proliferation of MDA-MB-231 breast cancer cells (Page 7164, right column, 2nd para.).
Regarding to claim 13, Fisher et al teach cancer cell lines were maintained in tissue culture flasks in an incubator, and all cell culture media were supplemented with 10% FBS (Page 7171, right column, 2nd para.).
Regarding to claim 14, Lutolf et al teach that the methods of the invention may be used to grow tumor-derived cells such as prostate tumors ([0134], page 10). FIG. 2D shows AG73-conjugated PEG matrices significantly enhance the growth of organoids ([0038], page 3). The hydrogel has a shear modulus between 0.05 -0.5 kPa ([0024], page 2). FIG. 10 shows generation of mouse and human tumor organoids in hydrogels of varying shear modulus and with or without RGD ([0046], page 4). Thus, a person of ordinary skill in the art would be able to use tumor-derived cells such as prostate tumors that grow ex vivo more slowly than their normal counterparts, with a shear modulus between 0.05 -0.5 kPa and with or without RGD.
Response to Arguments
Applicant's arguments filed on 10-29-2025 have been fully considered but they are not persuasive.
1. Applicant argue that the system taught by Lutolf et al is not suitable for freshly isolated or frozen human cells obtained from a human biopsy: “Gjorevski (Gjorevski et al., Designer matrices for intestinal stem cell and organoid culture, Nature, Vol 539, 24 November 2016, 560-56; Gjorevski et al., Synthesis and characterization of well-defined hydrogel matrices and their application to intestinal stem cell and organoid culture, Nature protocols, Vol. 12, no.11, 2017, 2263-2274; WO 2017/036533 A1 and WO 2017/037295 Al) ……. While some success of this approach was shown for the expansion and organoid formation from mouse cells, it was not shown (and rather questioned in Gjorevski 2017, p. 2265) that the above systems were suitable for the expansion and organoid formation from freshly isolated or frozen human cells from a biopsy of a human. Also, the only tested system, that is based on an enzymatic crosslinking reaction with factor XIII, is expensive, difficult to up-scale and/or to automatize for commercial purposes, and has also proven to be difficult to reproduce.” (Remarks, page 8-9)
Response to Arguments:
Applicant argues that the system taught by Lutolf et al is not suitable for use with freshly isolated or frozen human biopsy-derived cells, and emphasize that Lutolf allegedly demonstrated success only with mouse cell, relied on mouse-derived laminin, and disclosed systems that are expensive, difficult to scale , and difficult to reproduce. These arguments are not supported by the actual teachings of Lutolf et al because of the following reasons:
Lutolf et al teach PEG-based hydrogel as alternative to Matrigel: “owing to their chemically defined and versatile composition , synthetic ECM analogues (synthetic hydrogels , hereafter “hydrogels”) are considered attractive cell culture alternatives to natural, animal-derived matrices such as Matrigel …... It is known in the art that RGD functionalized PEG hydrogels with a low or high shear modulus are capable of supporting cell proliferation of cell lines” ([0007], page 1). Lutolf et al teach “the invention provides a method for epithelial tissue regeneration comprising a ) encapsulating and expanding of patient-derived epithelial stem cells or organoids in the three -dimensional hydrogel of the invention under suitable stem cell expansion conditions or suitable organoid formation conditions, and b ) transplanting the expanded stem cells or organoids back into the patient ([0031], page 2-3). Lutolf et al teach “The three-dimensional hydrogel system of the invention can be adapted for the culture of other types of adult epithelial stem cells and organoids, normal or tumor-derived” ([0066], page 6). Lutolf et al teach example 11: Growth of mouse and human tumor organoids: FIG.10 shows generation of mouse and human tumor organoids in hydrogels of varying shear modulus and with or without RGD . FIG . 10A :Mouse colon adenoma organoids in PEG -RGD matrices of varying stiffness, and quantification of colon adenoma organoid formation efficiency as a function of matrix stiffness. FIG . 10B : Human patient-derived colorectal cancer organoids grow efficiently in soft and stiff matrices , with or without RGD ([0181], page 15).
Additionally, Applicant’s arguments regarding cost, scalability, and difficulty of reproduction are not relevant to the determination of obviousness. Lutolf et al explicitly described these PEG hydrogel systems as defined and alternatives to matrigel, and obviousness does not require that prior art system be commercially optimized or clinically validated, only that it be workable for its intended use.
2. Applicant argue that “these deficiencies in art are generally discussed in the preamble of the present application. See page 10, line 25 to page 11, line 3 of the underlying PCT application (underlining added): Currently, the standard for the establishment of organoid cultures ex vivo includes firstly to encapsulate freshly isolated cells (from tissues) in the "gold standard" Matrigel …… Matrigel) is a gel derived from mouse sarcoma extract, which has already noted above has poor batch-to-batch consistency, has undefined composition and therefore cannot be used for clinical translational applications ……So far, there has not been a report of a successful expansion of freshly isolated or frozen human cells from biopsies or tissue resections …… There is thus also a need for providing a method for expansion of freshly isolated or frozen human cells from biopsies and subsequent formation of organoids therefrom, wherein said method completely avoids the use of a naturally-derived matrix such as Matrigel® and provides organoids suitable for clinical applications and generated in a commercially feasible manner, i.e. cost-effective, reliable, reproducible, automatable and up-scalable ……”
“Accordingly, even if Fischer et al. could be combined with Lutolf et al. as proposed by the Examiner (which is not conceded), Applicant submits the skilled person would not arrive at the subject matter of claim 1 amended herein (for the above reasons and for the reasons set out in responding to the rejection under 35 U.S.C. §102(a)(1 )). In fact, the skilled person would be lead away from any expectation of success adapting the teachings of Lutolf et al. to freshly isolated or frozen cells from a biopsy or tissue resection of a human or from patient-derived xenograft (PDX) tissue given the statement by the same author in Gjorevski 2017, p. 2265” (Remarks, pages 10-11).
Response to Arguments:
In response to applicant’s argument that there is no teaching, suggestion, or motivation to combine the references, the examiner recognizes that obviousness may be established by combining or modifying the teachings of the prior art to produce the claimed invention where there is some teaching, suggestion, or motivation to do so found either in the references themselves or in the knowledge generally available to one of ordinary skill in the art. See In re Fine, 837 F.2d 1071, 5 USPQ2d 1596 (Fed. Cir. 1988), In re Jones, 958 F.2d 347, 21 USPQ2d 1941 (Fed. Cir. 1992), and KSR International Co. v. Teleflex, Inc., 550 U.S. 398, 82 USPQ2d 1385 (2007). In this case, as described above, Lutolf et al explicitly described a fully defined three-dimensional hydrogel system comprising synthetic polymers such as PEG that is specifically designed for culturing and expanding both normal and tumor derived epithelial organoid by encapsulating and expanding cells under suitable conditions: Lutolf et al teach PEG-based hydrogel as alternative to Matrigel: owing to their chemically defined and versatile composition , synthetic ECM analogues (synthetic hydrogels , hereafter “hydrogels”) are considered attractive cell culture alternatives to natural, animal-derived matrices such as Matrigel ([0007], page 1). Lutolf et al teach “The three-dimensional hydrogel system of the invention can be adapted for the culture of other types of adult epithelial stem cells and organoids, normal or tumor-derived” ([0066], page 6). Lutolf et al also teach method of screening for activity of pharmacologic compounds or biomolecules on cells that are isolated from a biopsy sample (Claim 32, page 21). Lutolf et al also provide teaching for expanding of patient-derived epithelial stem cells or organoids in the three-dimensional hydrogel ([0031], page 2-3), and Lutolf et al teach example 11: Growth of mouse and human tumor organoids: FIG.10 shows generation of mouse and human tumor organoids in hydrogels of varying shear modulus and with or without RGD . FIG . 10A :Mouse colon adenoma organoids in PEG -RGD matrices of varying stiffness, and quantification of colon adenoma organoid formation efficiency as a function of matrix stiffness. FIG . 10B : Human patient-derived colorectal cancer organoids grow efficiently in soft and stiff matrices , with or without RGD ([0181], page 15).
Accordingly, the cited references show not only that tunable PEG hydrogels can support organoid and stem cell growth, but that such systems are expressly described for use with patient-derived biopsy samples in pharmacologic screening contexts, and are characterized as defined, reproducible and alternatives to matrigel. Applicant’s arguments concerning the unsustainability of Lutolf et al for human biopsy cells and practical limitations of the system are therefore not persuasive and do not overcome the rejections.
New-Claim Rejections - 35 USC § 102- necessitated by amendments with newly added claims
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 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.
Claims 21-22 are rejected under 35 U.S.C. 102 (a)(1) as being anticipated by Fisher et al (Adv. Funct. Mater. 2015, 25, 7163–7172, DOI: 10.1002/adfm.201502778).
Regarding to claim 21-22, Fisher et al teach tuning the microenvironment: click-crosslinked hyaluronic acid-based hydrogels provide a platform for studying breast cancer cell invasion (title) (For claim 1, the preamble).
Fisher et al teach 2×104 cancer cells per well were seeded on the hydrogel surface in 8 well chamber slides (Page 7171, right column, 3rd para.). The hyaluronic acid (HA)-based hydrogels crosslinked with matrix-metalloproteinase (MMP)-cleavable peptides are developed to study MDA-MB-231 breast cancer cell invasion. Hydrogels are synthesized by reacting furan-modified HA with bismaleimide peptide crosslinkers in a Diels–Alder click reaction. Increased crosslink density correlates with decreased MDAMB-231 cell invasion whereas incorporation of MMP-cleavable sequences within the peptide crosslinker enhances invasion (Abstract) (For claim 21, step a).
Fisher et al teach that the cells were cultured on the hydrogels for 4d before being fixed with 4% paraformaldehyde and stained with DAPI for visualization of cell nuclei (Page 7171, right column, 3rd para.) (For claim 21, step b).
Fisher et al teach determining the effects of tuning the hydrogels’ crosslink density, proteolytic degradation of the crosslinker, and ligand density on the invasion of MDA-MB-231 cells by hydrogel modifications: 1) To vary crosslink density, 1.25% w/v HA/MMPx hydrogels were formed from HA with varying furan substitutions. 2) To determine the influence of proteolytic degradation of the peptide crosslinker on invasion, medium crosslink density, 1.00% w/v HA hydrogels were formed with either MMPx or GAGx. 3) To confirm the influence of MMP-dependent invasion, MDAMB-231 cells were cultured on 1.25% w/v HA/MMPx with (25 × 10-6 M) or without (0 × 10-6 M) the MMP inhibitor GM6001. Both treatment groups contained 1% v/v DMSO in media. Hydrogels were preincubated in media containing 0 × 10-6 M or 25 × 10-6 M GM6001, and media were changed daily throughout the experiment. 4) To vary ligand density, prior to neutralizing HA/MMPx, pendant mal-GRGDS was dissolved in MES buffer and soaked into medium crosslink density, HA/MMPx hydrogels overnight (Page 7171, right column, 4th para.). It is noted that: As described above, Fisher et al teach culturing MDAMB-231 cells in different media with or without MMP inhibitor to have different treatment groups of MDAMB-231 cells; therefore, Fisher et al teach “repeating step b)” of growing and expanding cells in the presence of one or more different culture media (For claim 21, step c, and the claimed: wherein a specific combination of hydrogel features has been pre-selected for the said one tissue type to be tested and claim 22).
Thus, claims 21-22 are anticipated by Fisher et al.
Claims 23-25 are rejected under 35 U.S.C. 103 as being unpatentable over Fisher et al (Adv. Funct. Mater. 2015, 25, 7163–7172, DOI: 10.1002/adfm.201502778) in view of Lutolf et al (Pub. No.: US 2018/0258403 A1, Pub. Date : Sep.13, 2018).
The teachings of Fisher et al above are incorporated herein in their entirety.
Although Fisher et al teach cell screening to study cancer cell invasion, Fisher et al do not teach: a normal/healthy cell (claim 23); cells from a biopsy (claim 24); pancreatic ductal adenocarcinoma and PEG hydrogel having a stiffness in the range of 50 to 3000 Pa (claim 25); Lutolf et al who teach three dimensional hydrogels for culturing organoids (title) cure the deficiency.
Regarding to claim 23, Lutolf et al teach “Another aspect of the invention provides a method for culturing and expanding normal (healthy) epithelial organoids” ([0101], page 8)
Regarding to claim 24, Lutolf et al teach cells are isolated from a biopsy sample, and the cells are cultured in the presence or absence of the pharmacologic compounds or biomolecules to be tested (Page 21, claim 32).
Regarding to claim 25, Lutolf et al teach FIG. 9A: Mouse pancreatic ducts embedded in PEG -RGD ([0045], page 4) and the tumor-derived single cells can be derived from pancreatic tumors ([0104], page 8). Also, FIG.7A shows colony formation efficiency of ISCs embedded in degradable (DG) or non-degradable (N-DG) PEG gels of varying stiffness ([0043], page 4), and the three-dimensional hydrogels of the invention have an initial shear modulus (stiffness) of 0.5-5 kPa, preferably 0.5-2.5 kPa and a final shear modulus (stiffness) of 50-200 Pa ([0086], page 7). Additionally, FIG.9 shows culture and expansion of various mouse and human organoids in PEG-RGD ([0045], page 4). Thus, a person of ordinary skill in the art would be able to use non-degradable (N-DG) PEG gels of varying stiffness of 0.5-5 kPa or 50-200 Pa with a RGD motif.
Therefore, it would have been prima facie obvious for a person of ordinary skill in the art before the effective filing date of the rejected claims to combine the teachings of prior art to modify the method of Fisher et al by using a method for screening of libraries of pharmacologic compounds with three dimensional hydrogels for culturing organoids as taught by Lutolf et al as instantly claimed, with a reasonable expectation of success. Said modification amounting to combining prior art elements according to known methods to yield predictable results. One of ordinary skill in the art would have been motivated to do so because Lutolf et al provide several advantages such as AG73-conjugated PEG matrices significantly enhance the growth of organoids ([0038], page 3); Human patient-derived colorectal cancer organoids grow efficiently in soft and stiff matrices, with or without RGD ([0046], page 4); Laminin alone effectively supported colony formation in soft matrices ([0167], page 13). One of ordinary skill in the art would have had a reasonable expectation of success in doing so because Lutolf et al were successfully generated functionalized hydrogels for various cell types for screening of libraries of pharmacologic compounds, with detailed instructions, working examples and data.
Conclusion
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.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to KHOA NHAT TRAN whose telephone number is (571)270-0201. The examiner can normally be reached M-F (9-5).
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/KHOA NHAT TRAN/Examiner, Art Unit 1632
/PETER PARAS JR/Supervisory Patent Examiner, Art Unit 1632