DETAILED ACTION
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. Applicant’s Request for Continued Examination, Amendment and Arguments/Remarks received on 13 February 2026 have been entered. Claims 35, 38-39, and 45-47 were previously pending in the application. Claim 39 has been cancelled, and no new claims have been added by Applicant. Claim 35 is an independent claim.
Claims 35, 38, and 45-47 are currently pending and under examination in the instant application. An action on the merits follows.
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA . The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
Priority
The present application is a 35 U.S.C. 371 national stage filing of International Application No. PCT/US2020/020837, filed 03 March 2020, which claims priority to 62/813,307, filed 04 March 2019.
Thus, the earliest possible priority for the instant application is 04 March 2019.
Information Disclosure Statement
The information disclosure statement filed 13 February 2026 has been considered by the Examiner. Examiner notes the filing of IDS Size Fee assertions for the IDS filed 13 February 2026, as required under 37 CFR 1.98, indicating that no IDS size fee is required under 37 CFR 1.17(v) at this time.
Drawings
The drawings are objected to because Figure 8 presents a plot showing that Oncopig samples cluster by cell type based on transcriptomic profiles, as reproduced below. The figure legend indicates 7 cell/sample types presented in the plot using grey dots with slight intensity variation. However, the subtle differences in the intensity of the various grey dots do not allow for complete discernment of all cell/sample types within the plot.
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Appropriate correction is required.
Corrected drawing sheets in compliance with 37 CFR 1.121(d) are required in reply to the Office action to avoid abandonment of the application. Any amended replacement drawing sheet should include all of the figures appearing on the immediate prior version of the sheet, even if only one figure is being amended. The figure or figure number of an amended drawing should not be labeled as “amended.” If a drawing figure is to be canceled, the appropriate figure must be removed from the replacement sheet, and where necessary, the remaining figures must be renumbered and appropriate changes made to the brief description of the several views of the drawings for consistency. Additional replacement sheets may be necessary to show the renumbering of the remaining figures. Each drawing sheet submitted after the filing date of an application must be labeled in the top margin as either “Replacement Sheet” or “New Sheet” pursuant to 37 CFR 1.121(d). If the changes are not accepted by the examiner, the applicant will be notified and informed of any required corrective action in the next Office action. The objection to the drawings will not be held in abeyance.
Claim Rejections - 35 USC § 112(b)
The rejection of amended claim 47 under 35 U.S.C. 112(b) as failing to particularly point out and distinctly claim the subject matter which the inventor(s) regards as the invention for reciting “one or more driver or passenger mutations”, and “wherein the control comprises untreated mutated HCC cells or standard-of-care treatment”, and for being unclear how the method of claim 47 will identify a “personalized cancer treatment” by screening mutated porcine HCC cells which comprise additional cancer driver mutations (e.g., KRASG12D and TP53R167H) in addition to the one or more driver and/or passenger mutations which were identified by sequencing the patient’s HCC tumor cells and inserted into the HCC cells to produce the mutated HCC cells, is withdrawn in view of Applicant’s amendments to the claim.
The rejection of amended claim 46 under 35 U.S.C. 112(b) as failing to particularly point out and distinctly claim the subject matter which the inventor(s) regards as the invention for reciting “if cell growth and survival is affected by said compound compared to a control” is withdrawn in view of Applicant’s amendments to the claim defining the control as comprising HCC cells from the pig not treated with the compound.
Amended claims 35, 38, and 45-47 are newly 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. Claims 45-46 are included in this rejection due to their dependence on amended claim 38.
Amended claim 35 recites, “a pig comprising Cre recombinase-inducible porcine transgenes” in lines 1-2 such that Applicant removed the term “transgenic” from modifying the pig, which is indefinite because it is unclear whether Applicant intends for the pig to be comprise stable transgenes, such as transgenes integrated into the genome of the pig, which are comprised within all cells of the pig, or whether Applicant intends to encompass a pig comprising Cre recombinase-inducible porcine transgenes in a transient administration and/or within only a subset of cells within the pig.
As such, the metes and bounds of the claim cannot be determined.
Amended claim 38 recites, “producing cancer cells from the pig, wherein the cancer cells comprise hepatocellular carcinoma (HCC) cells”, which is indefinite because it is unclear how HCC cells are to be produced from the pig. For example, it is unclear whether the cancer cells are produced in vivo, ex vivo, or in vitro. It is also unclear whether the KRASG12D and TP53R167H transgenes are required to be activated by Cre recombinase in the process of producing the HCC cells; and if so, it is unclear whether the activation is occurring in vitro, ex vivo, or in vivo, in which cells the transgenes are activated, and how the Cre recombinase-dependent activation is being carried out. Therefore, it is unclear what active method steps are required to meet the limitation of “producing cancer cells”.
As such, the metes and bounds of the claim cannot be determined.
Amended claim 47 has multiple issues of indefiniteness.
Amended claim 47 recites, “developing HCC in the pig” in line 8, which is indefinite because it is unclear how HCC is being developed in the pig. For example, it is unclear whether the fibrosis induced within the pig, as recited in claim 35 is meant to lead to the development of the HCC within the pig over time without additional active method steps, including without activation of the KRASG12D and TP53R167H transgenes; or, alternatively, whether “developing HCC in the pig” is meant to encompass additional active method steps. If the latter, it is unclear whether the KRASG12D and TP53R167H transgenes are required to be activated by Cre recombinase in the process of developing the HCC, in which cells they need to be activated, and how the activation is meant to be done. It is also unclear whether the HCC is developed fully in vivo within the pig or whether the developing is meant to comprise additional ex vivo/ in vitro handling of cells (and if so, which cells and which handling steps) to promote the HCC development in the pig. Therefore, it is unclear what active method steps are required to meet the limitation of “developing HCC in the pig”.
Secondly, it is unclear whether Applicant intends for “the pig” recited on lines 9-10 to be the pig before or after developing HCC in the pig. It is also unclear whether Applicant intends for “the pig” recited on line 11 to have undergone the developing and testing steps prior to the determining step.
Thirdly, “determining a response to the one or more therapeutic interventions in the pig by comparing to a control” is indefinite because the sentence as written does not indicate what is being compared. The claim as written indicates that the response is determined by comparing to a control, but does not indicate what is being compared to determine the response. For example, it is unclear whether Applicant intends to compare some measurable trait relevant to the growth and/or survival of the HCC is which is assessed for the test pig and compared to a comparable growth and/or survival of the HCC trait assessed in the control pig, wherein the response is a relative relationship between the test and control measurements which would indicate the effectiveness and/or safety of the intervention being tested.
Fourthly, amended claim 47 recites, “with the same condition as the pig treated with the one or more therapeutic interventions” in each of lines 13-14 and 14-15, which is indefinite because it is unclear what is encompassed by “the same condition”. For example, it is unclear whether the pig merely has to have any HCC, whether the pig must have the KRASG12D and TP53R167H transgenes, whether the pig must be a pig with induced severe fibrosis according to claim 35, or whether the pig must be a second pig in which HCC was developed as recited in line 8 such that the pig is identical to the first pig in which HCC was developed according to line 8 and which was subsequently treated with one or more therapeutic interventions according to lines 9-10, wherein the second pig has the same Cre-inducible KRASG12D and TP53R167H transgenes and has undergone all the same “developing HCC” steps as the first pig.
As such, the metes and bounds of the claim cannot be determined.
Claim Rejections - 35 USC § 112(a)- Enablement
The rejection of amended claims 38 and 45-47 under 35 U.S.C. 112(a) for not being enabled for the full scope of the claims as written is withdrawn.
Amended claims 35, 38, and 45-47 are newly rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the enablement requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to enable one skilled in the art to which it pertains, or with which it is most nearly connected, to make and/or use the invention.
The claims are drawn to “a method of enhancing duration and severity of liver cirrhosis in a pig comprising Cre recombinase-inducible porcine transgenes KRASG12D and TP53R167H, in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery of the pig”, as recited in lines 1-4 of amended independent claim 35.
Claim 35 further recites a step of feeding the pig dietary ethanol comprising about 40% of total caloric intake for about four to eight weeks to produce more sustained and more severe liver fibrosis compared to ethanol infusion alone. Note that claim 35 as written does not require the pig to be fully transgenic for the Cre-inducible KRASG12D and TP53R167H transgenes present in all cells of the pig, the Cre-induced expression of the KRASG12D and TP53R167H transgenes, or the actual development of liver cirrhosis. Note additionally that the preamble requires that the pig be a pig “in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery of the pig” such that the active method step of feeding the pig dietary ethanol must occur following the induction of liver fibrosis by infusing ethanol into the common hepatic artery of the pig.
Amended claim 38 depends on claim 35 and recites “further comprising producing cancer cells from the pig, wherein the cancer cells comprise hepatocellular carcinoma (HCC) cells”. Note that although the step of producing cancer cells can occur before, after, or concurrent with the step of feeding the pig dietary ethanol, the preamble of claim 35 requires that the pig be a pig “in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery of the pig” such that the additional method step recited in claim 38 must occur following the induction of liver fibrosis by infusing ethanol into the common hepatic artery of the pig.
Amended claim 45 recites, “further comprising modeling one or more therapies for cancer treatment”, and amended claim 46 further limits the modeling to comprise “screening a potential therapeutic compound by contacting HCC cells from the pig with the compound to determine if cell growth and survival of the HCC cells is affected by the compound compared to a control, wherein the control comprises HCC cells from the pig not treated with the compound.” Note that although the step of modeling one or more therapies for cancer treatment can occur before, after, or concurrent with the steps of feeding the pig dietary ethanol and producing cancer cells from the pig, the preamble requires that the pig be a pig “in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery of the pig” such that the additional method step recited in claim 45 must occur following the induction of liver fibrosis by infusing ethanol into the common hepatic artery of the pig.
Amended claim 47 depends from claim 35 and recites, “further comprising developing HCC in the pig, testing one or more therapeutic interventions for HCC on the pig; and determining a response to the one or more therapeutic interventions in the pig by comparing to a control, wherein the control comprises i) an untreated pig with the same condition as the pig treated with the one or more therapeutic interventions, or ii) a pig treated with a standard-of-care HCC treatment with the same condition as the pig treated with the one or more therapeutic interventions”.
Note that although the steps of developing HCC in the pig, testing one or more therapeutic intervention for HCC on the pig, and determining a response to the one or more therapeutic interventions in the pig can occur before, after, or concurrent with the step of feeding the pig dietary ethanol, the preamble of claim 35 requires that the pig be a pig “in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery of the pig” such that the additional method steps recited in claim 47 must occur following the induction of liver fibrosis by infusing ethanol into the common hepatic artery of the pig.
Therefore, the scope of the claims encompasses any method of producing cancer cells comprising HCC cells from the pig without requiring a) the expression of the KRASG12D and TP53R167H transgenes, b) that the cells be hepatocytes originating in and isolated from the recited pig, or b) any active method steps which would facilitate the production of the HCC cells from the pig having the induced more sustained and more severe liver fibrosis.
The scope of the claims additionally encompasses any method of developing HCC in the pig without requiring the expression of KRASG12D and TP53R167H transgenes, without requiring a) that the HCC developed within the pig originate from hepatocytes of that pig, or b) any active method steps which would facilitate the development of the HCC within the pig having the induced more sustained and more severe liver fibrosis.
Note additionally that the term “Oncopig” is used herein to refer to a transgenic pig comprising Cre-inducible KRASG12D and TP53R167H transgenes.
This rejection comprises two (2) separate issues: 1) the absence of an enabling disclosure for producing cancer cells from a pig in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery, wherein the cancer cells comprise HCC cells; and 2) the absence of an enabling disclosure for developing HCC in a pig in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery.
These issues were identified by the Office after analysis of the disclosure provided by the specification. The Office has analyzed the specification in direct accordance to the factors outlined in In re Wands, namely 1) the nature of the invention, 2) the state of the prior art, 3) the predictability of the art, 4) the amount of direction or guidance present, and 5) the presence or absence of working examples, and presented detailed scientific reasons supported by publications from the prior art for the finding of a lack of enablement for the scope of the instant methods. The Wands analysis and supporting specific evidence are presented below for each of the identified issues.
As a first issue (1), the specification does not provide an enabling disclosure for producing cancer cells from a pig in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery, wherein the cancer cells comprise HCC cells.
The specification discloses “In other examples, metastatic liver disease is produced through in vivo autochthonous development of primary tumors, followed by biopsy, dissociation to develop cell lines, followed by autologous injection to produce metastatic tumors” [page 15 lines 7-9]. Therefore, the specification teaches that cancer cells can be produced from primary tumors, but does not teach how the autochthonous primary tumors are developed nor that they are specifically HCC cancer cells.
Additionally, Example 3 teaches, “As the OCM was designed to develop site- and cell-specific tumors in a temporal and spatial manner, additional tumors and cell lines representative of a variety of human cancers have been developed. Tumors developed to date in vivo include tumors reflective of human pancreatic ductal adenocarcinoma, soft-tissue sarcomas (STS), and HCC. The OCM cancer cell lines isolated and transformed in vitro are: fibroblasts, hepatocytes, pancreatic ductal cells, dermal epithelial cells, splenocytes, ovarian surface epithelial cells, Fallopian tube secretory epithelial cells, renal proximal tubule epithelial cells, bone marrow (no specific cell isolation), testis (no specific cell isolation), skeletal muscle (no specific cell isolation), and bladder epithelial cells.” [page 34 lines 6-13]. The specification also teaches that “Each of the listed cell lines has been isolated, transformed, shown to express the transgenes and form a tumor in SCID mice” [page 34 lines 15-16 and Table 7].
Therefore, the specification teaches that hepatocyte-derived cancer cell lines were generated from an Oncopig Cancer Model (OCM) pig, which comprises the Cre-inducible KRASG12D and TP53R167H transgenes, by isolation and transformation in vitro and that the hepatocyte-derived cancer cell lines express transgenes. However, the specification does not disclose how the cells were transformed, which transgenes are expressed, or whether the hepatocyte-derived cancer cell lines generated comprise HCC cells. To the extent that the transgenes expressed are the Cre-inducible KRASG12D and TP53R167H transgenes, the specification does not teach how they were induced for expression.
The specification also teaches generally that “Cells from the Oncopig can be transformed in culture with an adenovirus encoding Cre (AdCre)” [page 11 lines 9-10] and that “The pig cells may be tumor cells (such as Oncopig cells exposed to Cre recombinase to induce transgene expression) or noncancerous (e.g., primary) cells” [page 25 lines 16-18, see also page 40 lines 24-25]. Therefore, the specification teaches the requirement for Cre-recombinase induced transgene expression to produce cancer cells. However, the specification does not teach any actual examples of treating Oncopig cells with AdCre to transform the cells, the AdCre treatment of hepatocytes to specifically produce Oncopig HCC cells, nor the AdCre treatment of hepatocytes from Oncopigs in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery to specifically produce Oncopig HCC cells from a pig having liver fibrosis.
Example 7 teaches the use of Oncopig HCC cells [page 36 lines 23-27, page 37 lines 2, 8-9, 20-29], but does not teach the production of the HCC cell lines nor the use of a pig in which fibrosis has been induced. Likewise, Example 8 discloses use of HCC cell lines without teaching the production of the HCC cell lines nor the use of a pig in which fibrosis has been induced [page 38 lines 20-29, Table 10]. Example 10 provides prophetic protocols for using Oncopig cells for personalized tumor modeling, with a reference to the Oncopig HCC cells, but similarly does not teach the production of HCC cell lines from the pig nor the use of a pig in which fibrosis has been induced [page 40 lines 21-25, 31]. Further, the specification does not teach production of any cancer cells from any pig which has had fibrosis induced by infusing ethanol into the common hepatic artery of the pig.
Therefore, even though the specification generally teaches some general steps involved in producing cancer cells from an Oncopig, including general teachings of isolating cells and transforming cells by treating with AdCre, the specification does not teach a combination of steps to specifically produce cancer cells comprising HCC cells from a pig which has had fibrosis induced by infusing ethanol into the common hepatic artery of the pig. The specification does not provide any data indicating that applying the general teachings for isolating and transforming cells would work when applied to hepatocytes from an Oncopig in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery of the pig.
The art at the time of filing teaches that multiple factors, including viruses, chemicals, and inborn and acquired metabolic diseases are responsible for the development of HCC [Singh et al. 2018, Current Chemical Genomics and Translational Medicine, 12, 9-26, abstract]. Singh further teaches that HCC is the outcome of many variable etiological factors such as HBV, HCV, alcohol, aflatoxins, inborn and acquired metabolic disease, wherein the carcinoma might originate in mature liver cells or progenitor cells, and so the molecular basis of HCC progression may differ depending on diverse factors [page 3 section 4]. Singh further teaches that the development of HCC includes the onset of chronic hepatitis secondary to the factors described above, along with oncogene activation, which then leads to cirrhosis, resistance to cell death, loss of cell cycle control, and varied signaling pathway disruptions enroute to the onset of HCC [Figure 4, Table 2]. Therefore, the ordinarily skilled artisan would not expect a spontaneous production of cancer cells or development of cancer within a pig in the absence of risk factors or inducers such as HBV, HCV, alcohol, aflatoxins, inborn and acquired metabolic disease, and/or oncogene activation.
The art at the time of filing teaches that producing HCC cell lines from an Oncopig, which comprise the Cre-inducible KRASG12D and TP53R167H transgenes, requires the specific isolation of primary hepatocytes from the pig (pPH), followed by transformation of the isolated primary hepatocytes using AdCre in vitro to induce the expression of mutant KRASG12D and TP53R167H transgenes to produce pHCC as follows: “On day 2 of culture, the medium for each of the pPH cell lines was changed to low serum (5% FBS) and Ad5CMVCre-eGFP (AdCre; University of Iowa Vector Core, Iowa City, IA, USA) was added at a 200 to 500 MOI as previously described [4].” [Schachtschneider et al. 2017, Oncotarget, Vol. 8(38), 63620-63634, IDS, cited in a prior action, column 4 ¶ 2, column 19 ¶ 5-column 20 ¶ 2, Figure 1]. The art at the time of filing does not teach the production of HCC cancer cells from a pig without Cre-recombinase treatment nor from any pig in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery of the pig.
As such, neither the specification nor the art at the time of filing teaches producing cancer cells from a pig in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery of the pig. Additionally, neither the specification nor the art at the time of filing teaches producing cancer cells comprising HCC cells from an Oncopig without the expression of the KRASG12D and TP53R167H transgenes within the cells, without the cells being hepatocytes originating in and isolated from the Oncopig pig, nor without any active method steps which would facilitate the production of the HCC cells from the pig in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery of the pig.
Thus, in view of the art-recognized complexity of HCC development requiring various disruptions, including oncogene activation; the art-recognized requirement to isolate noncancerous cells from an Oncopig and then transform them by AdCre administration to induce KRASG12D and TP53R167H transgene expression in vitro to produce cancer cells from an Oncopig comprising Cre-inducible KRASG12D and TP53R167H transgenes, the lack of guidance in the specification for methods of isolating and transforming any cells to produce cancer cells comprising HCC cells from any Oncopig in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery of the pig, and the breadth of the claims, the skilled artisan would have considered producing cancer cells from the pig of claim 35, wherein the cancer cells comprise HCC cells, as highly unpredictable.
As a second issue (2), the specification does not provide an enabling disclosure for developing HCC in a pig in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery.
The specification discloses “In some examples, cell lines representative of human metastatic liver diseases are autologously injected into Oncopigs (e.g., subcutaneously or intrahepatically) to produce tumors. In other examples, metastatic liver disease is produced through in vivo autochthonous development of primary tumors, followed by biopsy, dissociation to develop cell lines, followed by autologous injection to produce metastatic tumors” [page 15 lines 7-9]. Therefore, the specification teaches that cell lines are autologously injected into Oncopigs either subcutaneously or intrahepatically to produce tumors, but does not teach which cells lines are injected nor the steps involved in obtaining and producing them. Additionally, the specification teaches that primary tumors can be produced in an Oncopig, but does not teach how the autochthonous primary tumors are developed nor that they are specifically HCC.
Example 2 of the specification discloses intrahepatic HCC tumor development in a cirrhotic liver environment, wherein the tumor developed in a pig in which cirrhosis was induced by infusion of ethanol into the common hepatic artery [page 27 lines 2-20, page 33 line 15- page 34 line 2, Figures 6A-6C]. Example 2 discloses the method for developing HCC in the pig as follows:
“Two weeks post autologous SQ tumor induction, SQ tumors were excised and dissected into roughly 1 mm segments. Utilizing a 15-gauge needle, tumor segments were inserted into the liver under ultrasound guidance. Intrahepatic tumor development was monitored every two weeks via ultrasound, and via CT 4-6 weeks post-intrahepatic tumor engraftment.” [page 33 lines 18-22].
However, Example 2 does not indicate a) how the “autologous SQ tumor induction” was carried out; b) what chemicals or cells were injected subcutaneously to induce the SQ tumor; c) if cells were injected subcutaneously, the source of the cells; nor d) whether the engraftment was performed before or after induction of cirrhosis within the liver.
Example 7 teaches genetically modified Oncopig HCC cell lines which were autologously injected subcutaneously into an Oncopig, wherein a tumor mass developed within two weeks; the subcutaneous mass was collected for histological and genomic analysis and determined to be an HCC tumor [page 37 lines 28-32]. However, Example 7 does not teach the source nor methods of producing the HCC cell lines. Additionally, Example 7 does not teach wherein the subcutaneous HCC tumor was produced in an Oncopig in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery nor without the additional genetic modification of the Oncopig cells.
Example 3 generally teaches that tumors have previously been developed in an OCM pig in vivo which are reflective of human HCC [page 34 line 9]. Example 10 presents a prophetic protocol for an in vivo model wherein mutations are introduced into Oncopig cells to render them tumorigenic, resulting in tumor formation [page 40 lines 33-35]. Neither Example 3 nor Example 10 teaches methods for developing HCC within any Oncopig, and specifically do not teach methods for developing HCC in an Oncopig in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery.
Accordingly, even though the specification generally teaches that it is possible to generate a primary tumor in an Oncopig, and further teaches some general steps involved in developing HCC in an Oncopig, including general teachings of injecting autologous cells either subcutaneously or intrahepatically, the specification does not teach a complete combination of steps to specifically develop HCC in any Oncopig nor specifically in an Oncopig which has had fibrosis induced by infusing ethanol into the common hepatic artery of the pig which would enable an ordinarily skilled artisan to perform the claimed method. The specification does not provide any data indicating that applying the general teachings for isolating cells, transforming cells, and injecting cells either subcutaneously or intrahepatically would result in developing HCC in an Oncopig in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery of the pig. The specification additionally does not teach the development of HCC in a pig, in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery of the pig, for testing one or more therapeutic interventions for HCC on the pig.
The art at the time of filing teaches that pPH cells which were specifically isolated from an Oncopig, transformed in vitro by treatment with AdCre to induce expression of the Cre-inducible KRASG12D and TP53R167H transgenes, and autologously injected subcutaneously into the same Oncopig yielded a tumor histologically characterized as HCC [Schachtschneider et al. 2017, Oncotarget, Vol. 8(38), 63620-63634, IDS, cited in a prior action, abstract, column 4 ¶ 2, column 6 ¶ 2, column 19 ¶ 5- column 20 ¶ 2, column 21 ¶ 5- column 22 ¶ 1, Figure 4]. Schachtschneider further teaches that to stage HCC with cirrhosis to define mono- or combination therapeutic approaches in a more controlled way than can be done in human patients requires further studies to produce orthotopic tumors via engraftment of subcutaneous HCC tumors into the liver of the Oncopig in which liver cirrhosis has been induced to form intrahepatic tumors [column 16 ¶ 1].
Schachtschneider additionally teaches autochthonous HCC tumors have been developed in a previously described chemically induced, non-Oncopig porcine HCC model, which takes over 1 year to develop clinically relevant tumors and does not allow for control of tumor number, location, or comorbidities, rendering it potentially less suitable for preclinical and co-clinical trials [column 16 ¶ 1- column 17 ¶ 1]. Therefore, given the teachings of Schachtschneider of a need for an induction mechanism to produce autochthonous HCC tumors, an ordinarily skilled artisan at the time of filing the instant application would have considered developing an autochthonous tumor in a pig model without any induction process as highly unpredictable.
As described above, the art at the time of filing also teaches that the development of HCC includes the onset of chronic hepatitis secondary to the factors described above, along with oncogene activation, which then leads to cirrhosis, resistance to cell death, loss of cell cycle control, chromosomal aberrations, and varied signaling pathway disruptions enroute to the onset of HCC [Figure 4, Table 1, 2]. Therefore, the ordinarily skilled artisan would not expect a spontaneous production of cancer cells or development of cancer within a pig in the absence of risk factors or inducers such as HBV, HCV, alcohol, aflatoxins, inborn and acquired metabolic disease, oncogene activation, chromosomal aberrations, and/or signaling pathway alterations.
Neither the specification nor the art at the time of filing teaches any method for developing in vivo autochthonous development of primary tumors in any Oncopig.
The specification and/or the art at the time of filing teach varied, general steps for producing HCC cancer cells lines and also teach the use of cancer cell lines for developing an autologous HCC in an Oncopig, wherein a combination of the varied methods comprise the specific method steps of a) specifically isolating hepatocytes from the Oncopig, wherein the cells are hepatocytes originating in and isolated from the same Oncopig pig; b) inducing expression of the KRASG12D and TP53R167H transgenes within the hepatocytes using Cre-recombinase; c) active method steps which would facilitate the production of the HCC cells from the pig; d) the subcutaneous injection of the autologous Oncopig HCC cells into the pig; e) the harvesting of the subcutaneous tumor from the Oncopig; and f) the subsequent autologous intrahepatic injection of a fragment(s) of the subcutaneous tumor within the pig. However, neither the specification nor the art at the time of filing teach to use the combination of steps disparately disclosed in a method of developing HCC in an Oncopig in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery of the pig. Further, neither the specification nor the art at the time of filing provide data that the combination of steps disparately disclosed would result in developing HCC in an Oncopig in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery of the pig.
As such, neither the specification nor the art at the time of filing teaches a sufficiently complete and predictable method for developing HCC in an Oncopig in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery of the pig for testing one or more therapeutic interventions for HCC. Additionally, as described by Singh, the art at the time of filing teaches that additional alterations are necessary for the development of HCC following development of cirrhosis [Figure 4]. As such, the ordinarily skilled artisan would consider spontaneous development of HCC in a pig lacking risk factors such as oncogene activation as highly unpredictable.
Thus, in view of the art-recognized unpredictability in developing an autochthonous tumor in a pig model without an induction mechanism; the art recognized unpredictability for producing cancer cells from a pig without isolation of specific cell types and transformation of the specific cell types to produce specific cancer cell types; the art recognized requirement to produce intrahepatic tumors for testing one or more therapeutic interventions for HCC; the art recognized requirement to isolate noncancerous cells from an Oncopig and then transform them by AdCre administration to induce KRASG12D and TP53R167H transgene expression in vitro to produce cancer cells from an Oncopig comprising Cre-inducible KRASG12D and TP53R167H transgenes; the lack of guidance in the specification for methods of isolating and transforming any cells to produce cancer cells comprising HCC cells from any Oncopig in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery of the pig; the further lack of guidance in the specification of methods for the generation of autologous subcutaneous tumors in an Oncopig to use for subsequent intrahepatic engraftment to generate intrahepatic tumors; and the breadth of the claims, an ordinarily skilled artisan would have considered developing HCC in the pig of claim 35 for testing one or more therapeutic intervention for HCC, as highly unpredictable.
As such, it would have required undue experimentation to produce cancer cells from an Oncopig in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery, wherein the cancer cells comprise HCC cells, and/or to develop HCC in an Oncopig in which liver fibrosis has been induced by infusing ethanol into the common hepatic artery for testing one or more therapeutic intervention for HCC, as encompassed by the claims.
Claim Rejections - 35 USC § 103
The rejection of amended and cancelled claims 35, 38-39, and 45-47 under 35 U.S.C. 103 as being unpatentable over Schachtschneider et al. [2017, Oncotarget, Vol. 8, 38, 63620-63634, IDS]; in view of Schuppan & Afdhal [2008, Lancet, 371, 838-851]; Halsted et al. [1993, Hepatology, Vol. 18(4), 954-960]; Halsted et al. [1996, Hepatology, Vol. 23(3), 497-505]; Constandinou et al. [2005, Methods in Molecular Medicine, 117, 237-250]; Schachtschneider et al. [2017, Frontiers in Oncology, Vol. 7, 190, 1-18, IDS]; Segatto et al. [2017, Frontiers in Pharmacology, 8, 894, 1-8, IDS]; Watson et al. [2016, Frontiers in Genetics, 7, 78, 1-16]; and Jones et al. [2015, Science Translational Medicine, Vol. 7(283), 1-10], is withdrawn in view of Applicant’s amendments to the claims and arguments of unexpected results, wherein the increasing severity of liver fibrosis at 4 weeks and 8 weeks of ethanol feeding at 40% of total caloric intake following induction of cirrhosis via hepatic artery infusion of ethanol compared to infusion alone is an unexpectedly high severity compared to an expectation gleaned from the cited prior, wherein motivation provided by the cited art suggested continued feeding to prevent recovery following the induction of cirrhosis via hepatic artery infusion of ethanol rather than to promote further 17278439increases in liver fibrosis severity.
Conclusion
No claim is allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to Dr. KATIE L PENNINGTON whose telephone number is (703)756-4622. The examiner can normally be reached M-Th 8:30 am - 5:30 pm, Friday 8:30 am - 12:30 pm CT.
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DR. KATIE L. PENNINGTON
Examiner
Art Unit 1634
/KATIE L PENNINGTON/Examiner, Art Unit 1634
Dr. A.M.S. Wehbé
/ANNE MARIE S WEHBE/Primary Examiner, Art Unit 1634
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