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 .
Continued Examination Under 37 CFR 1.114
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on 10/23/2025 has been entered.
Application Status
This action is written in response to applicant’s correspondence received on 10/23/2025. Claims 1-18 and 20-21 are pending. Claims 1 and 20 have been amended. Claim 19 has been cancelled. Claims 14-18 have been withdrawn. Claims 1-13 and 20-21 are currently under examination.
Claim Rejections - 35 USC § 112 – New Rejection Necessitated by Amendment
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 1-13 and 20-21 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 independent claims 1 and 20, these claims recite the term “-kine.” The term “-kine” is not a recognized terminology for a molecule such as a cytokine, and is therefore unclear. The specification offers some guidance surrounding the term “-kine” in paragraph 35, where hepatokines, adipokines, and cytokines are discussed. However, while the term “-kine” is recited in paragraph 35 after reference to these molecules, the term is not clearly defined. For instance, hepatocytes can secrete hepatokines and cytokines. It is unclear if the term “-kine” is meant to include only cell-specific secretions (e.g., hepatokines) or also general cytokines. It is recommended that the term “-kine” be replaced with a more descriptive and art-recognized term such as cytokine or adipokine, depending upon what the Applicant is attempting to claim.
Furthermore, regarding claim 12, claim 12 depends from claim 1 and recites “a hepatokine, an adipokine, and a cytokine crosstalk.” It is unclear if these newly recited “kines” are meant to be the same “kines” recited in claim 1 or if they are meant to be newly recited kines, in which case the additional recitation of claim 12 of for instance “the cytokine” would lack proper antecedent basis.
Claims 2-13 depend from claim 1, claim 21 depends from claim 20, and do not resolve these 112(b) issues and are therefore also rejected.
Further regarding claims 1 and 20, these claims recite the phrase “desired -kine concentration,” which is subjective terminology. Recitation of “desired -kine concentration” renders the claim indefinite because a “desired” concentration would change based upon an individual practitioners own subjective interpretation of the term, where the concentration can not be established based upon the present claim language.
Claims 2-13 depend from claim 1, claim 21 depends from claim 20, and do not resolve these 112(b) issues and are therefore also rejected.
Furthermore, regarding claim 20, claim 20 recites “kine concentration corresponding to each of the liver tissue cytoarchitecture in the chamber.” This claim language is confusing because only one liver tissue cytoarchitecture is recited in the claim. The phrase “each of the liver tissue cytoarchitecture in the chamber” makes it sound as if there are multiple liver tissue cytoarchitectures. It is therefore unclear how many liver cytoarchitectures are meant to recited.
Claim 21 depends from claim 20 and does not resolve this 112(b) issue and is therefore also rejected.
Claim Rejections - 35 USC § 103 – New Rejection Necessitated by Amendment
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.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claims 1-3, 7-9, 11-13, and 20-21 are rejected under 35 U.S.C. 103 as being unpatentable over Greene Nguyen (WO 2018/187380 A1, of record) in view of Ribeiro (Ribeiro AJS et al. Clin Pharmacol Ther. 2019 Jul;106(1):139-147, of record), Li (Li X et al. Lab Chip. 2018 Aug 21;18(17):2614-2631, of record), Wang (Wang YI et al. Adv Healthc Mater. 2018 Jan;7(2):10.1002/adhm.201701000, of record), Busek (Busek et al. J. Sens. Sens. Syst., 5, 221–228, 2016, of record), Tonon (Tonon F et al. Sci Rep. 2019 Sep 19;9(1):13557, of record), and Maas (Maass C et al. Integr Biol (Camb). 2017 Apr 18;9(4):290-302)
Regarding claim 1, Greene Nguyen is a patent document which focuses on the development of engineered liver tissue constructs which can be used to model liver disorders and test/develop therapeutics (Abstract, and see document). Greene Nguyen teaches a method for developing stratified medicine for nonalcoholic fatty liver disease, NAFLD: "in further embodiments, arrays for drug screening assays or drug discovery assays are used to research or develop drugs potentially useful in any therapeutic area," and "the liver disorder is a NAFLD," (paragraph 219 and 10, respectively. Greene Nguyen further teaches that:
"the liver disorder models offers a safe, economical, and clinically relevant platform to aid in the discovery of novel therapeutics to treat the condition. These liver disorder models can also be used for the identification of liver-specific biomarkers that can be easily measured noninvasively to diagnose and assess the severity of NAFLD," (paragraph 440)
Green Nguyen teaches a method that comprises obtaining a first microphysiological system, MPS, comprising a liver tissue cytoarchitecture:
"the present invention relates to a three-dimensional, engineered, bioprinted biological model of a liver disorder, comprising: a three-dimensional, engineered, bioprinted, biological liver tissue construct," (paragraph 6).
Green Nguyen further teaches:
"FIG. 18 is a series of non-limiting examples of planar and laminar geometries, including combinations thereof that are compatible with the methods of construction described herein, and reproduce architectural or spatial elements of native tissue architecture and biology," (paragraph 79).
Green Nguyen therefore teaches liver tissue cytoarchitecture in a first chamber in their methods (paragraphs 6 and 79).
Green Nguyen teaches obtaining a second MPS comprising an adipose tissue cytoarchitecture in a second chamber:
"following differentiation of pre-adipocytes to mature adipocytes, adipocytes can be cultured in the receiver wells of a 24-well plate with liver tissues cultured in the 24-well transwell basket," (paragraph 324)
and:
"FIG. 18 is a series of non-limiting examples of planar and laminar geometries, including combinations thereof that are compatible with the methods of construction described herein, and reproduce architectural or spatial elements of native tissue architecture and biology," (paragraph 79).
Green Nguyen teaches that the first MPS and the second MPS each comprise a compartment in a common fluidic system:
"following differentiation of pre-adipocytes to mature adipocytes, adipocytes can be cultured in the receiver wells of a 24-well plate with liver tissues cultured in the 24-well transwell basket," (paragraph 324)
Green Nguyen teaches that crosstalk occurs between the first MPS and the second MPS chambers based on fluid flow in the common fluidic system:
"the liver tissue is connected to one or more additional tissues constructs or cells via a fluid path or common fluid reservoir” (paragraph 222).
Green Nguyen teaches inducing metabolic dysfunction representing NAFLD in each of the first MPS and the second MPS chambers:
"the liver disorder (e.g., a NAFLD) is induced by applying at least two agents (i.e., at least two inducing agents) to the liver tissue construct selected from: a fructose, a glucose, a fatty acid, a lipopolysaccharide (LPS), a llip, a TNFalpha, or any combination thereof," (paragraph 274).
Green Nguyen further teaches that:
"similarly, the inducing agent can be any inducing agent, as described throughout this present disclosure, that is capable of inducing a liver disorder or phenotype thereof, such as NAFLD, NASH, a lipid accumulation, an inflammation, an oxidative stress, a fibrosis, a hepatocellular ballooning, a microvesicular steatosis, a macrovesicular steatosis, or any combination thereof,” (paragraph 351).
Green Nguyen teaches generating, based on inducing the metabolic dysfunction, transcriptomics data for each of the first MPS and the second MPS:
"bioprinted human liver tissues were treated for 14 days with either TGF-b to elicit a tissue-specific response (e.g., fibrosis) or a vehicle control. RNA was collected and used for microarray analysis according to standard procedures. Figure 46(A) shows differences observed in the expression profile of exemplary disease-associated genes by bioprinted human liver tissues after treatment with TGF-b ("TGF") as compared to a vehicle control ("VEH"), indicating biomarkers associated with tissue specific responses induced by TGF-b," (paragraph 435).
Green Nguyen further teaches that:
"the steps of evaluating the plurality of first and second media for genetic information comprises determining RNA levels in the plurality of first and second media by performing RNA sequencing on RNAs from the plurality of first and second media," (paragraph 368).
Green Nguyen teaches applying a drug to the first MPS and the second MPS using a dosing regimen: "dose-response experiments were conducted at doses typically ranging from 0.1 to 100 ng/ml, dependent on the ED50 of the experimental cytokine," (paragraph 392) and further teaches:
"thus, to build a robust liver disorder model for drug efficacy testing, a technical challenge is keeping the living cells viable and functional over a sufficient period of time (i.e., 3, 7, 14, 21, 28, 35 days) to cover (i) the initial time period needed to induce the disorder in the liver tissue construct, and (ii) the post-induction time period to evaluate the drug efficacy and/or toxicity of a candidate therapeutic agent on the liver disorder,” (paragraph 109).
Greene Nguyen further teaches monitoring changes in the transcriptomics data based on applying the drug:
"these liver disorder models can also be used for the identification of liver-specific biomarkers that can be easily measured noninvasively to diagnose and assess the severity of NAFLD," (paragraph 440).
and:
"bioprinted human liver tissues were treated for 14 days with either TGF-b to elicit a tissue-specific response (e.g., fibrosis) or a vehicle control. RNA was collected and used for microarray analysis according to standard procedures. Figure 46(A) shows differences observed in the expression profile of exemplary disease-associated genes by bioprinted human liver tissues after treatment with TGF-b ("TGF") as compared to a vehicle control ("VEH"), indicating biomarkers associated with tissue specific responses induced by TGF-b," (paragraph 435).
Thus, the models taught by Greene Nguyen relate the changes in the transcriptomics data to the dosing regimen of the drug (paragraph 435, above).
Regarding claim 2, Greene Nguyen teaches that the drug comprises one or more of small and large molecules configured to modulate activity of disease-relevant signaling pathways:
"the candidate therapeutic agent is selected from: a FXR agonist, a PPAR agonist, an ASK1 inhibitor, a GLP-1 agonist, a DPPIV inhibitor, an AMPK activator, a mTOR inhibitor, a 11B-HSD1 inhibitor, a FGF19 analogue, a FGF19 analogue, an anti-mRNA, a caspase inhibitor, a SCD1/ACC inhibitor, a LXRalpha inhibitor, a DGATI inhibitor, a vitamin E, a vitamin E analogue, a leptin receptor agonist, a statin, a cholesterol absorption inhibitor, an iBAT inhibitor, a KHK inhibitor, a galectin-3 inhibitor, a broad spectrum immunomodulator, an antioxidant, a TNFa inhibitor, a PDE inhibitor, an ATI receptor agonist, a CCR2/CCR5 antagonist, a TLR4 antagonist, a LPS antibody, a L TD4 receptor antagonist, an inflammasome modulator, a LOXL2 immunotherapy, a HSP47 inhibitor, a TGFP inhibitor, or any combination thereof,” (paragraph 353).
Regarding claim 3, Greene Nguyen teaches that: “the top panel of Figure 40 shows that NASH-induced (12.5 mM fructose and 500 uM PA) bioprinted liver tissue constructs treated with the FXR agonist, Obeticholic Acid (OCA),” (paragraph 429).
Regarding claim 7, Greene Nguyen teaches inducing metabolic dysfunction representing NAFLD in each of the first MPS and the second MPS comprises one or more of: inducing one or more of insulin resistance, IR, excessive de nova
gluconeogenesis and lipogenesis, and dysregulated hepatokine signaling in the first MPS:
"Cytokine stimulation was conducted by adding cytokine directly to the culture media and incubating the bioprinted tissues with the added protein to provide direct and prolonged cell access to the proper stimulus" (paragraph 392).
Green Nguyen teaches inducing one or more of IR, increased lipolysis, and dysregulated adipokine signaling in the second MPS:
"the liver tissue construct is induced to exhibit at least one phenotype of a NAFLD by contacting the liver tissue construct with two or more inducing agents selected from: a fructose, a fatty acid a lipopolysaccharide (LPS), a llip, a TNFa, or any combination thereof,” (claim 81).
Regarding claim 8, Greene Nguyen teaches that inducing the metabolic dysfunction comprises: characterizing one or more of phenotypic, biomarker, and transcriptomic signatures of NAFLD pathology; and determining, a physiological relevance of one or more phenotypes, biomarkers, or transcriptomics of NAFLD:
"methods of identifying a biomarker associated with at least one phenotype characteristic of a liver disorder, the method further comprises determining whether the biomarker is differentially expressed in liver tissue in a subject having a liver disorder as compared to expression of the biological molecule in liver tissue in a subject without the liver disorder,'' (paragraph 35)
and "the liver tissue construct is induced to exhibit at least one phenotype of a NAFLD,” (claim 81).
Regarding claim 9, Greene Nguyen teaches applying the model to one or more stratified patient subpopulations based on differential biological mechanisms for each stratified patient subpopulation:
"the method comprising the steps of: receiving a plurality of media from a three-dimensional, engineered, bioprinted liver tissue construct exhibiting only a non-alcoholic fatty liver disease (NAFLD); receiving a plurality of serum from a patient with a comparable NAFLD and at least one other liver disorder; evaluating the plurality of media from the liver tissue construct for genetic information; evaluating the plurality of serum from the patient with the comparable NAFLD for genetic information; and identifying one or more biomarkers that differentiate the evaluated genetic information of the liver tissue construct from the evaluated genetic information of the patient," (paragraph 371).
Regarding claim 11, Greene Nguyen teaches generating, based on applying a model, one or more of phenotypic, transcriptomic, and metabolomic datasets establishing a molecular characterization of each stratified patient subpopulation in:
"to compare the present invention's in-vitro liver disorder models to clinical data, a qualified medical professional, such as a pathologist, can score the in-vitro liver disorder tissues for steatosis, hepatocellular ballooning, and fibrosis using the NASH and fibrosis scoring system, then using available serum samples from NAFLD patients with comparable steatosis, hepatocellular ballooning, and fibrosis scoring, compare the histopathology, secreted proteome, and RNA sequencing profiles between our in vitro samples and clinical samples to potentially identify biomarkers that can delineate the various stages of NAFLD," (paragraph 363).
Regarding claim 12, Greene Nguyen teaches connecting the first MPS and the second MPS by milli -fluidic recirculation:
"the engineered liver tissues are subjected to continuous or periodic perfusion, recirculation, or agitation of liquid nutrients on one or more surfaces. In other embodiments, the engineered liver tissues and arrays thereof are housed in a multi-well bioreactor that provides continuous or periodic recirculation of the liquid culture media for each construct" (paragraph 157).
Green Nguyen teaches facilitating one or more of a hepatokine, an adipokine, and a cytokine crosstalk between the first MPS and the second MPS cytoarchitectures:
"the liver tissue is connected to one or more additional tissues constructs or cells via a fluid path or common fluid reservoir," (paragraph 222)
and:
"Cytokine stimulation was conducted by adding cytokine directly to the culture media and incubating the bioprinted tissues with the added protein to provide direct and prolonged cell access to the proper stimulus," (paragraph 392).
Greene Nguyen further teaches the scaling of each of the first MPS and the second MPS are scaled based on relative sizes of cytoarchitecture/cell populations human physiology for one or more of the hepatokine, the adipokine, and the cytokine crosstalk:
"physiologically relevant populations (e.g., hepatic stellate cells (hSC) and human aortic endothelial cells (EC)) of cells were combined at specific ratios to generate proper bio-ink," (paragraph 389)
and teaches "bioprinted neotissues containing physiologically -relevant populations of cells were successfully stimulated with cytokines that had been previously demonstrated to elicit cellular responses in two-dimensional in vitro systems," (paragraph 393).
Regarding claim 20, Greene Nguyen discloses a method for developing stratified medicine for nonalcoholic fatty liver disease, NAFLD:
"arrays for drug screening assays or drug discovery assays are used to research or develop drugs potentially useful in any therapeutic area," (paragraph 219).
Nguyen further teaches:
"the liver disorder is a NAFLD, wherein: (a) the liver tissue construct exhibits at least one phenotype selected from: a lipid accumulation, an inflammation, an oxidative stress, a fibrosis, a hepatocellular ballooning, a microvesicular steatosis, a macrovesicular steatosis, or any combination thereof,” (paragraph 10)
and further teaches:
"the liver disorder models offers a safe, economical, and clinically relevant platform to aid in the discovery of novel therapeutics to treat the condition. These liver disorder models can also be used for the identification of liver-specific biomarkers that can be easily measured noninvasively to diagnose and assess the severity of NAFLD," (paragraph 440).
The method of Greene Nguyen comprises: obtaining a microphysiological system, MPS, comprising a chamber comprising a liver tissue cytoarchitecture:
"the present invention relates to a three-dimensional, engineered, bioprinted biological model of a liver disorder, comprising: a three-dimensional, engineered, bioprinted, biological liver tissue construct," (paragraph 6)
and:
"FIG. 18 is a series of non-limiting examples of planar and laminar geometries, including combinations thereof that are compatible with the methods of construction described herein, and reproduce architectural or spatial elements of native tissue architecture and biology,” (paragraph 79).
Greene Nguyen further teaches inducing metabolic dysfunction representing NAFLD in the liver tissue of the MPS:
"the liver disorder (e.g., a NAFLD) is induced by applying at least two agents (i.e., at least two inducing agents) to the liver tissue construct selected from: a fructose, a glucose, a fatty acid, a lipopolysaccharide (LPS), a llip, a TNFalpha, or any combination thereof,” (paragraph 274)
Greene Nguyen teaches generating, based on inducing the metabolic dysfunction, transcriptomics data for the MPS:
"bioprinted human liver tissues were treated for 14 days with either TGF-b to elicit a tissue-specific response (e.g., fibrosis) or a vehicle control RNA was collected and used for microarray analysis according to standard procedures. Figure 46(A) shows differences observed in the expression profile of exemplary disease-associated genes by bioprinted human liver tissues after treatment with TGF-b ("TGF") as compared to a vehicle control ("VEH"), indicating biomarkers associated with tissue specific responses induced by TGF-b," (paragraph 435).
Greene Nguyen teaches applying a drug to the MPS using a dosing regimen:
"dose-response experiments were conducted at doses typically ranging from 0.1 to 100 ng/ml, dependent on the ED50 of the experimental cytokine," (paragraph 392)
and further:
“the post-induction time period to evaluate the drug efficacy and/or toxicity of a candidate therapeutic agent on the liver disorder," (paragraph 109).
Greene Nguyen teaches monitoring changes in the transcriptomics data based on applying the drug:
"these liver disorder models can also be used for the identification of liver-specific biomarkers that can be easily measured noninvasively to diagnose and assess the severity of NAFLD" and paragraph 435: "bioprinted human liver tissues were treated for 14 days with either TGF-b to elicit a tissue-specific response (e.g., fibrosis) or a vehicle control. RNA was collected and used for microarray analysis according to standard procedures. Figure 46(A) shows differences observed in the expression profile of exemplary disease-associated genes by bioprinted human liver tissues after treatment with TGF-b ("TGF") as compared to a vehicle control ('VEH"), indicating biomarkers associated with tissue specific responses induced by TGF-b,” (paragraph 440).
Greene Nguyen teaches generating a model relating the changes in the transcriptomics data to the dosing regimen of the drug:
"bioprinted human liver tissues were treated for 14 days with either TGF-b to elicit a tissue-specific response (e.g., fibrosis) or a vehicle control. RNA was collected and used for microarray analysis according to standard procedures. Figure 46(A) shows differences observed in the expression profile of exemplary disease-associated genes by bioprinted human liver tissues after treatment with TGF-b ("TGF") as compared to a vehicle control ('VEH"), indicating biomarkers associated with tissue specific responses induced by TGF-b," (paragraph 435)
and:
"the figure shows that protection against disease progression was achieved upon co-administration of OCA, providing proof of concept for application of the NASH model in drug efficacy assessments," (paragraph 429)
Regrading claim 21,Greene Nguyen teaches exposing tissues to shear forces in their systems as a result of fluid flow and recirculated fluid flow (paragraph 209). Furthermore, Greene Nguyen teaches the concept of protecting/minimizing cell damage caused by shear forces (paragraph 140).
Regarding claims 1 and 20 Greene Nguyen, while teaching separate chambers for liver and adipocyte cytoarchitectures, and that fluid crosstalk can be exchanged through chambers through recirculated culture media, does not teach that the chambers are connected by a pump to a reoxygenation chamber, where the reoxygenation chamber is configured to add oxygen to the fluid loop and mix the oxygen into the fluid loop (paragraphs 6, 324, 222, and paragraph 157). Green Nguyen does not teach that a first size of the first chamber and a second size of the second chamber (claim 1) and/or a cell culture surface area (claim 20) are selected to establish an oxygen-dependent liver metabolic zonation profile, the first size and second size (claim 1) or the surface area (claim 20) being scaled based on measured -kine secretion rates for the liver tissue cytoarchitecture and for the adipose tissue cytoarchitecture, the oxygen-dependent liver metabolic zonation profile representing desired -kine concentrations corresponding to each of the liver tissue cytoarchitecture in the first chamber and the adipose tissue cytoarchitecture in the second chamber.
Regarding claim 21, Greene Nguyen does not teach or suggest that the shear force is less than 0.05 dyne/centimeter or providing oxygen tension to the tissue cytoarchitecure.
Ribeiro is a review article that specifically focuses on liver microphysiological systems (MPS) for predicting and evaluating drug effects (Title, Abstract, and see document). The subject matter of Greene Nguyen and Ribeiro therefore directly overlaps because they are both in the same field of endeavor.
Regarding liver MPS systems, Ribeiro teaches that:
“different liver MPS (Figure 2d–f) can vary in the design of microfluidic devices, pumping systems, profile of media circulation (Figure 5), coculture of varied cell types, and their connectivity to other MPS that represent different organs,” (page 144, right column, final paragraph).
Furthermore, Ribeiro teaches that MPS systems for liver cells comprise a pump for recirculating media (Figure 5). Ribeiro therefore teaches that recirculating media through various chambers of liver MPS systems using a pump is known and is a part of the design of MPS systems (page 144, right column, final paragraph and Figure 5).
Additionally, Ribeiro teaches that:
“according to the US National Institutes of Health, they [MPS] are defined as microfluidic systems that enable the coculture of at least two types of human cells in three dimensions,” (page 139, left column, first paragraph).
Therefore, according to the definition taught by Ribeiro, the apparatus taught by Greene Nguyen is an MPS system because Greene Nguyen teaches recirculating fluidics and coculturing of two types of human cells in three dimensions (paragraphs 157, 6, and 324).
Ribeiro further teaches that:
“With regard to media flow, the importance of controlling oxygen concentration in MPS to mimic liver physiology as a function of oxygen tension is also described,” (page 141, right column, second paragraph).
Ribeiro therefore teaches that control of the influx/flux of oxygen concentration in MPS is important so that such MPS systems can properly mimic liver physiology (page 141, right column, second paragraph).
Ribeiro further teaches Figure 2 which teaches oxygen gradient formation/flow in liver MPS (Figure 2), and further teaches that:
“microfluidic chambers can also maintain tissue function, under oxygen gradients,” page 141, caption below Figure 2).
Ribeiro references Li with regards to the caption beneath Figure 2 (page 141, caption below Figure 2).
Li is a research article which focuses on liver MPS systems designed for modeling diseases (Title, Abstract, and see document). The subject matter and field of endeavor or Greene Nguyen, Ribeiro, and Li therefore directly overlaps. Furthermore, the fact that Ribeiro directly references Li is further proof that the two are in the same field.
Li teaches that:
“In hard plastic or glass sealed microfluidic devices, the sole source of oxygen for cells is the influx of cell culture media (~170–200 μM52, 53). In the vLAMPS, the hepatocytes and NPCs are fed by two independent flow channels, the vascular channel and the hepatic chamber,” (page 5, final paragraph).
Li therefore teaches multiple channels designed for the influx/flow from different cell groups in MPS systems, and further that this is a source of reoxygenation in MPS systems (page 5, final paragraph).
Furthermore, Li also teaches that:
“by utilizing microfabrication, bioprinting, or the combination of both, as well as sequential layering of cells, human liver MPS have been developed as promising alternative or complementary models,” (page 3, second paragraph).
Li therefore teaches that liver MPS have been developed as promising alternative models to bioprinting (page 3, second paragraph).
Wang is a review article that focuses on multi-organ MPS systems and their role in drug development (Title, Abstract, and see document). Wang therefore directly overlaps in field of endeavor and subject matter with Greene Nguyen, Ribeiro, and Li. Wang teaches embodiments where different cell types are connected via pump-driven recirculating platforms which comprise a pump with connecting fluid channels, where fluid is flowed by the pump between multiple chambers in a fluid loop part of a common fluidic system (e.g., Figure 2C). Wang teaches that interconnected channels allow for crosstalk between tissue types including between liver and fat/adipocyte tissues (page 4, first paragraph). Wang therefore teaches general methods of connecting MPS chambers, that such chambers can be connected to generate crosstalk between tissues, that such connections can be connected with a pump for fluid flow in a common fluidic loop, and furthermore that connected tissues that allow for crosstalk broaden the research capacity of such systems beyond those of traditional static cell culture models (page 4, first paragraph, Figure 2C). Wang therefore provides motivation to a practitioner to include fluidics which allow for crosstalk between tissue compartments, as well as teaching MPS designs that include pumps, recirculating media, and common fluidics/fluid loops (page 4, first paragraph, and Figure 2C).
Thus, the pump-driven recirculating MPS system taught by Wang appears to teach recirculating fluid flow by a pump between a first and second chamber with two different tissue types, a media reservoir, and crosstalk between the MPS based on the recirculating fluid flow in the common fluidic loop (Figure 2C). Further, Li teaches that media can be a source of reoxygenation (page 5, final paragraph).
In addition, Busek is a research article that teaches the design, characterization, and modelling of microcirculation systems with integrated oxygenators (Title, Abstract, and throughout). Busek teaches that several human tissue systems, including multiple-organ chips (MOCs), have been cultivated within such systems, including liver cells (Introduction, page 221, right column, first paragraph). Busek therefore directly overlaps in subject matter and field of endeavor with Green Nguyen, Ribeiro, Li, and Wang, because each document concerns the development of microcirculation systems comprising the same elements (e.g, multiple tissues/cell types, fluid circuits, microfluidics, pumping mechanisms, etc.). Furthermore, Busek teaches that oxygenator elements can be added as additional elements implemented in microfluidics platforms, allowing for automated and reproducible control of oxygen levels (Introduction, final paragraph). Busek teaches microfluidic systems comprising two cultivation chambers A and B for cell types, pumps, and oxygenator chambers (Figure 2b). Busek teaches that such systems are compatible with multi-organ modelling systems, including liver modeling (Introduction, final paragraph). Busek teaches that previous fluidic designs relied upon the pump acting as both a flow source and oxygenator (page 222, left column, second paragraph). Busek teaches that such designs can be problematic, and teaches the alternative design of pumping fluidics through an oxygenator (i.e., a reoxygenation chamber), where oxygen is transported (mixed) within the fluidic system (Figure 2b, page 222, left column second paragraph into right column first paragraph). Busek therefore teaches that there are known designs of reoxygenation chambers which can add and mix oxygen into the fluid loop (Figure 2b and page 222, left column second paragraph into right column first paragraph).
Additionally, in regards to claims 1 and 20 and also 13, Tonon is a research article focused on metabolic zonation (Abstract and throughout). Tonon teaches that:
“the aim of this study is to understand whether an oxygen gradient, generated in vitro in our developed device, is sufficient to instruct a functional metabolic zonation during the differentiation of human embryonic stem cells (hESCs) from endoderm toward terminally differentiated hepatocytes,” (Abstract).
Tonon therefore teaches human physiologies comprising oxygen-dependent liver metabolic zonation profiles, and furthermore taught that such oxygen-dependent metabolic zonation profiles have been not only taught but reduced to practice and applied to microphysiological systems (Abstract and throughout). Tonon further teaches that:
“the device developed can physiologically induce the differentiation of hepatocytes which, in the future, may be used to deepen our knowledge about the physiology/pathology of periportal and perivenous hepatocytes and possibly to reconstitute the diseased liver tissue,” (Conclusion, final paragraph).
Tonon teaches that:
“in standard differentiation cultures it is still problematic to accurately reproduce the phenomenon occurring in vivo with regard to the differentiation of stem cells towards hepatocyte-like cells. Moreover, hepatic differentiation from human pluripotent stem cells requires long-term experiments and up to 3 weeks could be needed for functional differentiation. Therefore, simple and easy to operate devices are desired from the biological community. The use of advanced technologies can minimize the above problems and better mimicking the in vivo differentiation process. Thus, advanced technologies have the potential to allow the generation of hepatocytes usable for drug screening, toxicological assays and may be also as liver disease in vitro models,” (Discussion, first paragraph).
Thus, Tonon teaches that their methods can be applied advantageously for drug screenings and toxicological assays to more accurately reproduce in vivo characteristics (Discussion, first paragraph).
Tonon teaches that chamber width (i.e., size) is known to affect metabolic zonation cell profiles (e.g., Figure 1B and 1C). Tonon therefore taught that chamber size was a known variable which can be adjusted to affect the outcome of oxygen-gradient dependent zonation (Figure 1 and caption). Furthermore, Tonon also teaches that it is known that cytokine gradients (i.e., concentrations) are known to have an effect in the induction of liver zonation profiles and are a consideration when designing such MPSs (Conclusion, second paragraph).
Regarding the teaching that the first size and second size are scaled based upon -kine secretion and desired -kine concentrations, Maass is a research article that focuses on the design of multi-functional MPS scaling and methodologies of designing such using various input parameters, where such multi-functional designing provides designs that are “better” than standard MPS scaling (Abstract, and throughout). Maass teaches that the multi-functional scaling method can be applied across MPS designs comprising liver cytoarchitectures in combination with other tissues (e.g., page 291, right column, second paragraph). Maass teaches that their scaling design algorithms can be applied to individual MPSs (claim 20) or multi-MPS platforms (claim 1) where such designs recapitulate functional characteristics of biological states (page 291, left column, fourth paragraph). Maass teaches that cytokine production rates and concentrations are such examples of such biological functions to be considered to be integrated into their parameters and designs for MPSs (page 291, left column, fourth paragraph). Maass further teaches and contemplates scaling specifically using cytokines (i.e., -kines) as an input value in their multi-functional scaling algorithm (page 298, right column, fifth paragraph). Furthermore Maass teaches motivational teachings to apply their methodology and scaling algorithm and designs across multiple designs, and that the design and use of their scaling is easily implementable and beneficial:
“The novel multi-functional scaling approach described here
allowed the design of integrated multi-MPS platforms for relevant
pharmacological applications. In the applications demonstrated
here, the resulting design parameter values are practical and can
be easily implemented. The approach can be readily adapted to
various multi-MPS platforms for a variety of study purposes,” (Conclusion).
Regarding claim 21, Li teaches supplying oxygen tension to tissues inside of MPS chambers (page 6, first paragraph and bottom of page 7). Li teaches that the shear force of the fluid flow on the tissues of their system is less than 0.05 dyne/square centimeter because they teach that the shear-stress in their systems is approximately 2.5 x 10-7 dyne/centimeter squared (page 24, caption beneath Figure 1). Li teaches that such shear stress levels are well within a range that would not negatively affect liver cell functionality and health (page 13, first paragraph).
It would have been obvious to a person of ordinary skill in the art before the effective filing date of the claimed invention to modify the teachings of Greene Nguyen with those of Ribeiro, Li, Wang, Busek, Tonon, and Maass to arrive at a multichannel liver MPS system with a pump and reoxygenation chamber where the reoxygenation chamber can be configured to add and mix oxygen into the fluidic system/loop because such a combination is the simple combination of known prior art elements. In the present case, Ribeiro teaches that pumps are common components of MPS systems, and further that control of oxygen is critical for maintaining relevant physiological conditions in liver MPS systems (page 144, right column, final paragraph and page 141, right column, second paragraph, respectively). Li teaches multi-channel liver MPS systems where oxygen influx is controlled through culture media circulation (page 5, final paragraph). Furthermore, Wang teaches embodiments of MPS systems that include multiple chambers that are connected by common fluidic loops, which are controlled by a pump and recirculate microfluids, and that such systems allow broaden the capacity for research by allowing crosstalk between tissue chambers (page 5 third paragraph, page 4 first paragraph, Figure 2C). Busek teaches that oxygenation chambers/oxygenator chambers which can both add and mix oxygen into a fluid circuit are known and have been reduced to practice (Abstract, Figure 2b). A practitioner would therefore be motivated to combine the teachings of Greene Nguyen, Ribeiro, Li, and Wang in order to properly mimic liver physiology in liver MPS systems. This is particularly true in light of the fact that the art appears to be replete with teachings concerning the claim elements recited, where multi-organ tissue/cell systems are known to be models for NAFLD, and furthermore where reoxygenator chambers are also known, as well as the importance of maintaining oxygen levels and gradients in order to properly model such tissue systems (e.g., Riberio page 141, right column, second paragraph). Thus, a practitioner would be motivated to apply the teachings of Busek, who teaches such reoxygenation strategies of introducing and mixing oxygen in a chamber to supply to the fluid loop in multi-organ/cell tissue microfluidics systems.
Additionally, a practitioner would be motivated to use scaling such as that taught by Maass in multi-functional MPS systems, where scaling of the MPS chambers is based upon input values such as cytokine concentration and secretion in established oxygen-dependent metabolic zonation profiles because Tonon teaches the establishment of oxygen metabolic zonation profiles, that such states have been reduced to practice to recapitulate more accurately in vivo conditions, and furthermore that cytokine concentrations are known to affect such zonation profiles (Tonon, above). Furthermore, Maass teaches the known method of scaling based upon inputs such as cytokines, which are known to be relevant for metabolic zonation profiles per Tonon. Furthermore, a practitioner would be motivated to combine the teachings of Tonon and Maass with Green Nguyen, Ribeiro, Li, Wang, and Busek, as such methods of Tonon and Maass recapitulate more accurately in vivo physiological modeling (per Maass). The results are further predictable because Maass teaches that such scaling is easily implementable across MPS platforms and provides data which correlate highly with in vivo scenarios.
Additionally, Li teaches that their shear stress levels, which are under 0.05 dyne/centimeter, do not adversely affect liver cells, and so would be an obvious range of shear stress level given that Greene Nguyen is designed to function with liver cells (Li, page 13, first paragraph, and caption of Figure 1).
With regard to the interpretation that Greene Nguyen teaches bioprinting as opposed to MPS systems, even if this were the case (which it is not because Greene Nguyen teaches fluidic recirculation of two separate human tissues which defines MPS according Ribeiro), Li teaches that MPS systems are promising alternatives to bioprinting for modeling (Ribeiro page 139, left column, first paragraph and Li page 3, second paragraph). A practitioner would therefore be motivated to adopt the bioprinting systems taught by Greene Nguyen to MPS systems because Li teaches that MPS systems are promising alternatives to bioprinting (Li, page 3, second paragraph).
Claims 4 and 5 are rejected under 35 U.S.C. 103 as being unpatentable over Greene Nguyen (WO 2018/187380 A1) in view of Ribeiro (Ribeiro AJS et al. Clin Pharmacol Ther. 2019 Jul;106(1):139-147), Li (Li X et al. Lab Chip. 2018 Aug 21;18(17):2614-2631), Wang (Wang YI et al. Adv Healthc Mater. 2018 Jan;7(2):10.1002/adhm.201701000), Busek (Busek et al. J. Sens. Sens. Syst., 5, 221–228, 2016) Tonon (Tonon F et al. Sci Rep. 2019 Sep 19;9(1):13557, of record), and Maas (Maass C et al. Integr Biol (Camb). 2017 Apr 18;9(4):290-302)
as applied to claims 1-3, 7-9, 11-13, and 20-21 above, and further in view of Chung (Chung JY et al. Genome Res. 2019 Sep;29(9):1442-1452).
A discussion of the teachings of Greene Nguyen, Ribeiro, Li, Wang/Busek, Tonon, and Maass regarding claims 1-3, 7-9, 11-13, and 20-21 is given above.
Regarding claim 4, Greene Nguyen in combination with Ribeiro, Li, and Wang renders obvious the method of claim 1, including applying a drug using a dosing regimen (paragraph 392 and see rejection of claim 1 above).
Regarding claim 5, Greene Nguyen teaches targeting FXR:
"the top panel of Figure 40 shows that NASH-induced (12.5 mM fructose and t,00 uM PA) bioprinted liver tissue constructs treated with the FXR agonist, Obeticholic Acid (OCA), exhibited a reduction in steatosis and fibrosis,” (paragraph 429).
Regarding claim 4, Greene Nguyen, Ribeiro, Li, and Wang do not teach or suggest that the drug comprises CRISPR short guide RNAs (sgRNAs) that modulate an activity or expression of disease-relevant signaling pathways.
Regarding claim 5, Greene Nguyen, Ribeiro, Li, and Wang while do not teach or suggest the use of sgRNAs to modulate expression.
Chung is a research article which focuses of the use of iCRISPR systems targeting Fabp4 in adipocytes to ameliorate obesity, inflammation, hepatic steatosis, and insulin resistance (Title, Abstract, and throughout). Chung teaches CRISPR short guide RNAs (sgRNAs) that modulate an activity or expression of disease-relevant signaling pathways:
"CRISPR interference (CRISPRi) mechanism based on catalytically dead Cas9 (dCas9) and single guide RNA (sgRNA) ... Targeted delivery of the CRISPRi system against Fabp4 to white adipocytes by ATS-9R induced effective silencing of Fabp4, resulting in reduction of body weight and inflammation and restoration of hepatic steatosis in obese mice. This RNA-guided DNA recognition platform provides a simple and safe approach to regress and treat obesity and obesity-induced metabolic syndromes," (Abstract).
Chung also teaches that:
"this condition, known as steatosis, leads to other hepatic morbidities like nonalcoholic fatty liver disease, cirrhosis, and steatohepatitis," (page 1446, col 1, paragraph 1).
Thus, the scope of Greene Nguyen, Ribeiro, Li, Wang, and Chung overlap because they concern liver diseases, drug treatments for liver diseases, and specifically NAFLD.
It would have been obvious to a person of ordinary skill in the art before the effective filing date of the claimed invention to modify the method of Greene Nguyen, Ribeiro, Li, Wang, Busek, Tonon, and Maass to include the CRISPR sgRNA drug taught by Chung because such a combination is the simple substitution of one known element (the drugs used by Greene Nguyen) for another (the sgRNA CRISPR drug taught by Chung) to obtain predictable results. Greene Nguyen teaches an array of drugs for screening and drug therapy discovery, and screening the CRISPR drug taught by Chung is therefore the simple substitution of one drug for another. The results are predictable because the drug treatments are for the same class of disease (liver disease, NAFLD). Furthermore, a practitioner would be motivated to test the drug taught by Chung, because Chung has taught that the drug is simple and safe to use, and thus a practitioner would be motivated to test such a drug’s efficacy in the models and methods taught by Greene Nguyen.
Regarding claim 5, as discussed above, Chung teaches iCRISPR as drugs to target liver conditions and modulate expression (Abstract).
It would have been obvious to a person of ordinary skill in the art before the effective filing date of the claimed invention to use the method of Greene Nguyen, Ribeiro, Li, Wang, Busek, Tonon, and Maass and to include Chung’s sgRNA to target FXR to modulate its expression because such a combination is the simple combination of prior art elements to yield predictable results. As discussed above, a practitioner would be motivated to test the CRISPR sgRNA drugs taught by Chung in their methods, simply to screen the efficacy of the drugs in their models. Furthermore, Greene Nguyen taught that targeting FXR exhibited a reduction in steatosis and fibrosis. Thus, FXRwould have been an obvious target of sgRNA drugs, taught by Chung.
Claim 6 is rejected under 35 U.S.C. 103 as being unpatentable over Greene Nguyen (WO 2018/187380 A1) in view of Ribeiro (Ribeiro AJS et al. Clin Pharmacol Ther. 2019 Jul;106(1):139-147), Li (Li X et al. Lab Chip. 2018 Aug 21;18(17):2614-2631), Wang (Wang YI et al. Adv Healthc Mater. 2018 Jan;7(2):10.1002/adhm.201701000), Busek (Busek et al. J. Sens. Sens. Syst., 5, 221–228, 2016) Tonon (Tonon F et al. Sci Rep. 2019 Sep 19;9(1):13557, of record), and Maas (Maass C et al. Integr Biol (Camb). 2017 Apr 18;9(4):290-302)
as applied to claims 1-3, 7-9, 11-13, and 20-21 above, and further in view of Bhise (Bhise NS et al. Biofabrication. 2016 Jan 12;8(1):014101). Claim 6 is further evidenced by Widjiati (Widjiati et al. HAYATI J. of Biosciences, 19:1, published March, 2012).
Regarding claim 6, the combination of Greene Nguyen, Ribeiro, Li, Wang/Busek, Tonon, and Maass, renders obvious the method of claim 1, including a dosing regimen comprising applying a drug at various concentrations (Greene Nguyen, paragraph 392 "Dose-response experiments were conducted at doses typically ranging from 0.1 to 100 ng/ml, dependent on the ED50 of the experimental cytokine"). Greene Nguyen further teaches in that they used TGF-β1 at concentrations of 0,1,10, and 50 ng/ml (paragraph 76). As evidenced by Widjiati, the molecular weight of TGF-β1 is 25 kDa (Abstract, first sentence). Thus, Greene Nguyen teaches a concentration that ranged from 0-2 nanomolar, and which establishes a lower boundary of drug regimen testing (the nanomolar range) in liver MPS systems.
Greene Nguyen, Ribeiro, Li, and Wang do not teach or suggest that the drug regimen involves applying the drug at five concentrations spanning nano-to-milli-molar ranges.
Bhise is a research article focused on using liver-on-a-chip platforms (i.e., liver microphysiological states) to evaluate drug testing regimen ranges and their effects on engineered hepatic constructs (Abstract). Bhise teaches that they tested drug concentrations with their synthetic liver constructs in the range from 0, 1, 5, 10, and 20 mM. Thus, Bhise teaches an upper limit of drug concentration testing in synthetic liver constructs of at least 20 mM.
Furthermore, the subject matter of Greene Nguyen, Ribeiro, Li, Wang, and Bhise directly overlaps because they relate to the construction of engineered hepatic tissue systems and testing drug therapies with these systems. Bhise also teaches that there is a pressing need to understand toxicity effects of drugs on liver tissue (Abstract). Thus, Bhise teaches a motive to test a range of drugs, to better understand potential toxic effects that drugs may have on liver tissue (Abstract).
It would have been obvious to a person of ordinary skill in the art before the effective filing date of the claimed invention to combine the teachings of Greene Nguyen, Ribeiro, Li, Wang, Busek, Tonon, and Maass and Bhise to arrive at the present invention because such a combination is the simple combination of known prior art elements to yield predictable results. Greene Nguyen taught a lower limit of drug regimen testing in the nanomolar range, while Bhise taught that higher concentrations of drugs (in the millimolar range) are also known in the art to be tested in engineered liver tissue systems. Thus, a practitioner would have a reasonable expectation of success when testing drugs across the dosing regimen ranges recited in the present claim because drugs at these concentrations have been reduced to practice in the art. Furthermore, a practitioner would be motivated to test a range of drugs in the recited liver systems of Greene Nguyen and Bhise to understand not only their efficacious ranges, but also their potential toxicity to liver tissues, as taught by Bhise (Abstract).
Claim 10 is rejected under 35 U.S.C. 103 as being unpatentable over Greene Nguyen (WO 2018/187380 A1) in view of Ribeiro (Ribeiro AJS et al. Clin Pharmacol Ther. 2019 Jul;106(1):139-147), Li (Li X et al. Lab Chip. 2018 Aug 21;18(17):2614-2631), Wang (Wang YI et al. Adv Healthc Mater. 2018 Jan;7(2):10.1002/adhm.201701000), Busek (Busek et al. J. Sens. Sens. Syst., 5, 221–228, 2016), Tonon (Tonon F et al. Sci Rep. 2019 Sep 19;9(1):13557, of record), and Maas (Maass C et al. Integr Biol (Camb). 2017 Apr 18;9(4):290-302) as applied to claims 1-3, 7-9, 11-13, and 20-21, above, and further in view of Kostrzewski (Kostrzewski T et al. 2019 Nov 13;4(1):77-91).
Regarding claim 10, the combination of Greene Nguyen, Ribeiro, Li, Wang, Busek, Tonon, and Maass renders obvious the methods of claims 1 and 9.
Greene Nguyen, Ribeiro, Li, and Wang do not teach or suggest that the differential biological mechanism comprises one of a high-risk genetic single nucleotide polymorphism or a gender-specific hormone.
Kostrzewski is a research article focused on microphysiological systems (MPS) for studying nonalcoholic steatohepatitis and NAFLD (Title, Abstract, and throughout). Kostrzewski teaches that:
“the genetic basis of NAFLD has started to be explored, and the I148M mutation in the patatin-llke phospholipase domain containing 3 (PNPLA3) gene has been identified as the major genetic variant that associates with NAFLD/NASH progression. This single nucleotide polymorphism (SNP) can lead to more than 2-fold increases in hepatic fat content in NAFLD patients,” (page 77, first paragraph).
Kostrzewski therefore teaches a biological mechanism which comprises one of a high-risk genetic single nucleotide polymorphism (SNP), (page 77, first paragraph).
It would have been obvious to a person of ordinary skill in the art before the effective filing date of the claimed invention to combine the teachings of Greene Nguyen, Ribeiro, Li, and Wang with Kostrzewski to use the models and methods taught by Greene Nguyen, Ribeiro, Li, and Wang with the teaching of Kostrzewski that the PNPLA3 gene SNP is a high risk biological mechanism for developing NAFLD because such a combination is the simple combination of known prior art elements to yield predictable results. Greene Nguyen taught arrays for drug screening and drug discovery, and Kostrzewski taught the I148M mutation in PNPLAS3 as a major genetic risk variant associated with NAFLD progression. A person of ordinary skill in the art would recognize that the high risk mutation taught by Kostrzewski could be combined with the drug discovery and therapeutic models taught and suggested by Greene Nguyen, Ribeiro, and Li for mutation-specific NAFLD disorders.
Response to Arguments
The Applicant’s arguments submitted 10/23/2025 have been considered but are not persuasive. The Applicant has made amendments to independent claims 1 and 20, and in effect argues that these the amendments render the recited invention non-obvious as the prior art does not teach the new claim elements. This argument is not found to be persuasive because the amendments prompted a new search which lead to the rejections set forth above. Briefly, the prior art teachings of Maass were applied to the rejection, where Maass teaches scaling and designing of MPS platforms using biologically functional input data such as cytokines, and that such design parameters and algorithms are easily implementable to multi-functional and single MPS systems, where furthermore such scaling designs offer advantages in that they reliably recapitulate vivo modeling using MPS. Furthermore, the previous prior art Tonon is incorporated into the rejection of the independent claims, where Tonon teaches and reduces to practice metabolic liver zonation in an MPS, and furthermore specifically teaches that cytokine gradients (i.e., concentrations) are critical factors when considering liver metabolic zonation profiles and cell characteristics (Conclusion, and see 103 rejection above). Thus, the prior art teaches that such oxygen-dependent liver metabolic zonation profiles have been reduced to practice in MPSs and furthermore teaches methods to and motivations to incorporate cytokine secretion and concentrations into designing such MPS (Tonon and Maass, above).
112(a) – New Rejection Necessitated by Amendment
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.
Claims 1-13 and 20-21 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.
Regarding independent claims 1 and 20, these claims recite MPS systems which have chambers (claim 1) or a cell surface area (claim 20), which have been scaled based upon cytokine secretion levels in the context of metabolic liver zonation profiles, where the size of the chamber or surface area is recited with a functional outcome (i.e., to establish an oxygen-dependent liver metabolic zonation with a desired -kine secretion). The claims are therefore claiming a structural-functional relationship between the size of the chambers and the effect of generating a functional outcome of establishing a liver metabolic zonation with any given desired profile based upon -kine secretion. This claim language is problematic because the Applicant has not established any such structure-function relationship in the drawings, specification, or claims sufficient to claim such a scope as what is claimed.
With regards to the guidance provided in the specification, as an initial matter, the Applicant’s remarks filed 10/23/2025 indicate that support for the amendments (i.e., the recited “sizes” of chambers or surface areas corresponding to zonation, where the sizes/surface area are scaled based on measured -kine secretion rates of tissue architecture) can be found in paragraphs 31-35, 68, and Figures 1A-1D. Paragraphs 31-34 describe an overview of MPS designs, which are depicted in Figures 1A-1D. Paragraph 35 recites:
“The compartment 122a-b geometries are estimated using a multi-functional scaling methodology. The geometries are optimized to establish an oxygen-dependent liver metabolic zonation profile which is relevant to NAFL-NASH disease progression. For example, a relationship between severity and zonal location of steatosis to the presence of NASH include that there is an increased disease severity (e.g., fibrosis and ballooning) in zone 3 (perivenous - low oxygen) compared to zone 1 (periportal- high oxygen). Briefly, for the multi-functional scaling of liver-adipose MPS 120, a computational model for the adipose-liver axis is focused on crosstalk molecules, such as hepatokines (liver-function), adipokines (adipose-function) and cytokines (both adipose and liver). The measured -kine secretion rates, subsequently described, are the inputs for these models, while additional literature data are used as either as input parameters (e.g. -kine receptor and Kd values) or the output functions (such as -kine plasma and/or portal vein concentrations, in vitro -kine dose response curves). A multi-functional scaling algorithm optimizes the relative tissue sizes (e.g., the input function) to fit a desired the -kine concentrations (e.g., the output function) at which a biological effect is observed.
Thus, paragraph 35 alludes to a scaling methodology where geometries are optimized and a multi-functional scaling algorithm to optimize relative tissue sizes (above) but does not describe such an algorithm or recite any geometries (i.e., chamber sizes or surface areas) associated with an observed functional output such as a desired metabolic zonation profile.
Paragraph 68 further elaborates upon MPS design principles, and states that: “[g]enerally, the integrated chip compartment sizes (cell culture surface area, seeding density and volume) are informed with the multi-functional mechanistic scaling methodology described previously… [t]he concentration gradient and distribution of molecules secreted from cells or molecules/drugs introduced in the system are
simulated,” (paragraph 68).
With regards to the specification and drawings in general, these do not appear to offer any specific chamber sizes or surface areas which would result in the recited oxygen-dependent liver metabolic zonation, where for instance a first size and second size have been scaled based on -kine secretion. The Applicant has therefore not shown possession of a structure with the recited required limitations, where furthermore the scaling/sizing of the chambers and surface areas are recited with a specific function. The specification appears to be prophetic with regards to these claim limitations but does not show that such scaling has been reduced to practice to have such a desired outcome. The Applicant was therefore not in possession of the recited chamber sizes which have been scaled to have the recited functional outcomes. The Applicant has not reduced to practice or characterized such a structure-function relationship.
Furthermore, with regards to the state-of-the art, Tonon (Tonon F et al. Sci Rep. 2019 Sep 19;9(1):13557, of record) is a research article focused on metabolic zonation through oxygen gradient on a chip (Title, Abstract, and throughout). Tonon teaches the establishment of oxygen-dependent metabolic zonation on a chip/MPS (Abstract, throughout). Tonon teaches that:
“we cannot exclude that other factors can be involved in the induction of liver zonation in our system. It is known that, in vivo, dynamic gradients of different modulators such as cytokines and/or signaling molecules contribute to the zonation effect,” (Conclusion, second paragraph)
Thus, Tonon teaches that in the context of zonation effects, dynamic gradients of cytokines can influence the induction of liver zonation in such systems. However, the instant application does not appear to have characterized any such effects or concentrations of cytokines or secreted cytokines with respect to how such a parameter would influence chamber size design, and was not in possession of MPS designs which are scaled based upon cytokine secretions and which are capable of oxygen-dependent liver metabolic zonation profiles with “desired” cytokine concentrations.
Claims 2-13 which depend from claim 1 and claim 21 which depends from claim 20 do not resolve the 112(a) issue and are also rejected.
In short, the Applicant did not show possession of scaled chamber sizes based upon cytokine secretion, a structural feature, because no such scaled chamber sizes/dimensions appear to be recited in the specification.
Conclusion
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/D.C.R./Examiner, Art Unit 1635
/KIMBERLY CHONG/Primary Examiner, Art Unit 1636