DETAILED ACTION
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Applicant’s Response to Election/Restriction Filed, Amendment, and Arguments/Remarks, filed 25 July 2025, have been entered. Claims 1-4 and 6-22 are currently pending. Claims 1 and 12 are independent claims. Applicant’s election of the following species:
Cells: d. patient-specific megakaryocyte progenitors,
Medicaments: a. Eltrombopag,
Gene variants: a. ANKRD26-RT,
Proteins of the extracellular matrix: e. fibronectin,
without traverse in a reply filed 25 July 2025 is acknowledged.
Claims 1-4 and 6-22 are currently pending in the application and under examination to which the following grounds of rejection are applicable. An action on the merits follows.
Priority
The present application is a 35 U.S.C. 371 national stage filing of International Application No. PCT/US2020/063639, filed 07 December 2020, which claims priority to U.S. Provisional Application No. 62/944,874, filed 06 December 2019.
Thus, the earliest possible priority for the instant application is 06 December 2019.
Information Disclosure Statement
The information disclosure statements filed 01 February 2024 and 23 July 2025 have been considered by the Examiner. Filing of a size fee assertion in accordance with 37 CFR 1.98 for the IDS filed 23 July 2025, indicating that no IDS size fee is required under 37 CFR 1.17(v) at this time, is acknowledged.
Specification
The use of the term “Triton X-100” in [0048], “Alexa Fluor” in [0048, 0050, 0052], “Hoescht” in [0048, 0050], “ProLong Gold” in [0048, 0052], “Olympus BX51” in [0048, 0052], “TCS SP8” in [0050], “GraphPad” in [0056], which is a trade name or a mark used in commerce, has been noted in this application. The term should be accompanied by the generic terminology; furthermore the term should be capitalized wherever it appears or, where appropriate, include a proper symbol indicating use in commerce such as ™, SM , or ® following the term.
Although the use of trade names and marks used in commerce (i.e., trademarks, service marks, certification marks, and collective marks) are permissible in patent applications, the proprietary nature of the marks should be respected and every effort made to prevent their use in any manner which might adversely affect their validity as commercial marks.
Claim Objections
Independent claims 1 and 12 are objected to because of the following informalities:
Claims 1 and 12 recite, “a) preparing from cells acquired from the patient and/or acquiring from the patient patient-specific haematopoietic stem cells” on lines 4-5 and 3-4, respectively, which appear to be missing a colon after the second recitation of “patient” so as to read , “a) preparing from cells acquired from the patient and/or acquiring from the patient: patient-specific haematopoietic stem cells”.
Claims 1 and 12 are written in an informal language style which lacks clauses such as “wherein” which serve to clarify the claim limitations. For example, use of the terms/phrases “with”, “including”, “includes”, “upon having”, and “incorporation” represent information language which promotes ambiguity. Further, the lack of terms such as “wherein” as an adverb for modifying clauses contributes to the ambiguity in that the relationship of the clauses to the prior limitations is unclear. For example, recitation of “f) comparing platelet production from the patient-specific megakaryocytes within the experimental model versus the patient-specific megakaryocytes within the control model, the control model lacking the medicament” is lacking a word to connect the latter phrase “the control model lacking the medicament” to the prior recitation of “comparing platelet production from the patient-specific megakaryocytes within the experimental model versus the patient-specific megakaryocytes within the control model”. Additionally, steps b), e), and g) likewise recite phrases offset by commas which lack connecting words which relate the phrases to the other limitations within the claim. Appropriate correction is required.
Claim 6 is objected to because of the following informalities: claim 6 recites the abbreviations “ANKRD26-RT” and “MYH9-RD” without first writing out the terms for which they are abbreviations. Appropriate correction is required.
Claim 17 is objected to because of the following informalities: claim 17 recites “claim 12wherein” which appears to be a typographical error for “claim 12, wherein”. Claim 17 additionally recites the abbreviations “ANKRD26-RT” and “MYH9-RD” without first writing out the terms for which they are abbreviations. Appropriate correction is required.
Claim 18 is objected to because of the following informalities: claim 17 recites “claim 12 the method” which appears to be a typographical error for “claim 12, the method”. Appropriate correction is required.
Claim Rejections - 35 USC § 112(b)
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 1-4 and 6-22 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.
Independent claims 1 and 12 each recite, “cells acquired from the patient and/or acquiring from the patient” in lines 4 and 3, respectively, which is indefinite because it is unclear in what manner the cells are acquired from the patient, such that it is unclear whether the patient-specific cells recited are specific to the same patient from which they are acquired, or whether the patient may be providing cells which are specific to a different patient. As such, the metes and bounds of the claim cannot be determined.
Independent claims 1 and 12 each recite, “patient-specific haematopoietic stem cells, patient-specific haematopoietic progenitor cells, patient-specific Induced Pluripotent Stem Cells, and/or patient-specific megakaryocyte progenitors” and subsequent lists referring specifically to the recites cells in lines 4-6, 7-9, 17-19, 21-23 in claim 1 and lines 3-5, 6-8, 16-18, 20-22 of claim 12. Currently, it is not clear which species are included in the group and which are not. It is unclear whether the group of “patient-specific haematopoietic stem cells, patient-specific haematopoietic progenitor cells, patient-specific Induced Pluripotent Stem Cells, and/or patient-specific megakaryocyte progenitors” being selected from is an open or closed group. As such, the metes and bounds of the claim cannot be determined. See MPEP § 2117.
Independent claims 1 and 12 additionally recite, “haematopoietic stem cells, haematopoietic progenitor cells, Induced Pluripotent Stem Cells, and/or megakaryocyte progenitors” in lines 13-14 of claim 1 and lines 12-13 of claim 12. Currently, it is not clear which species are included in the group and which are not. It is unclear whether the group of “haematopoietic stem cells, haematopoietic progenitor cells, Induced Pluripotent Stem Cells, and/or megakaryocyte progenitors” being selected from is an open or closed group. As such, the metes and bounds of the claim cannot be determined. See MPEP § 2117.
Independent claims 1 and 12 also recite, “patient-specific megakaryocytes, including cell adhesion, proplatelet extension, and platelet release and collection” in lines 23-24 and 22-23, respectively, which is indefinite because it is unclear whether the step of “maturing” is meant to include “cell adhesion, proplatelet extension, and platelet release and collection” or whether the “patient-specific megakaryocytes” are meant to include “cell adhesion, proplatelet extension, and platelet release and collection”. Further, if the step of “maturing” is meant to include “cell adhesion, proplatelet extension, and platelet release and collection”, then it is unclear what active steps are being claimed by “cell adhesion”, “proplatelet extension”, and “platelet release” as they are characteristics, activities carried out by the cells themselves. Additionally, if the “patient-specific megakaryocytes” are meant to include “cell adhesion, proplatelet extension, and platelet release and collection”, then it is unclear in what way the megakaryocytes include “collection”. Therefore, the metes and bounds of the claims cannot be determined.
Independent claim 1, line 17 of step c) recites “introducing the medicament to the patient-specific “. The preamble of claim 1 is directed to a method of administering a medicament wherein the medicament is indicated to modulate megakaryocyte differentiation”. However, steps a) to g) result in administration of the medicament if there is a prediction that exceeds a threshold supporting the recitation of the preamble. Thus the antecedent bases for “the medicament” in line 17 is unclear. Therefore, the metes and bounds of the claims cannot be determined.
Independent claims 1 and 12 also recite, “f) comparing platelet production from the patient-specific megakaryocytes within the experimental model versus the patient-specific megakaryocytes within the control model” in lines 25-27 and 24-26, respectively, which is indefinite because it is unclear how platelet production from cells within one model can be compared versus the cells themselves in a different model. Therefore, the metes and bounds of the claims cannot be determined.
Further, claims 1 and 12 recite informal language with terms that are imprecise and result in ambiguity in the claim language. Claims 1 and 12 recite the terms “including” and “includes” in lines 15, 23, and 28 of claim 1 and lines 14, 22, and 27 of claim 12, which is ambiguous because it is unclear if the terms “including” and “includes” indicates an open or a closed group.
Claims 1 and 12 recite the phrase “a prediction of in vivo platelet production of the patient upon having the medicament administered”, which is indefinite because it is unclear whether the prediction is predicting the results of medicament administration, whether the prediction is predicting the production at the time of administration of the medicament, or whether the prediction is being made at the time of administration of the medicament.
Also, claims 1 and 12 recite the term “with” in lines 31 and 30, respectively, which is ambiguous because it is unclear if the limitations following the term “with” are additional components of the system or whether they further limit the existing components.
Further, the lack of terms such as “wherein” as an adverb for modifying clauses contributes to the ambiguity in that the relationship of the clauses to the prior limitations is unclear. For example, recitation of “f) comparing platelet production from the patient-specific megakaryocytes within the experimental model versus the patient-specific megakaryocytes within the control model, the control model lacking the medicament” is lacking a word to connect the latter phrase “the control model lacking the medicament” to the prior recitation of “comparing platelet production from the patient-specific megakaryocytes within the experimental model versus the patient-specific megakaryocytes within the control model”.
Additionally, steps b), e), and g) likewise recite phrases offset by commas which lack connecting words which relate the phrases to the other limitations within the claim, e.g., “each of the at least two three-dimensional bone marrow models” in lines 9-10 (claim 1) and 8-9 (claim 12), “the at least two three-dimensional bone marrow models” in lines 14-15 (claim 1) and 13-14 (claim 12), “each of the regenerated silk fibroin sponges” in line 15 (claim 1) and 14 (claims 12), “including cell adhesion” in line 23 (claim 1) and 22 (claim 12), “the prediction incorporating” in lines 29 (claim 1) and 28 (claim 12), and “expressed either as fold increase or absolute platelet numbers” in line 31 (claim 1) and 30 (claim 12). Therefore, the metes and bounds of the claims cannot be determined.
Independent claims 1 and 12 recite, “wherein the prediction is validated”, which is indefinite because it is unclear whether the validation is an active step of the claimed method or whether the validation is merely modifying the structure of the prediction such that the prediction already has the attribute of a statistically significant correlation between an increase in platelet count ex vivo and in vivo prior to incorporating the comparing of step f). Further, if the latter, then it is unclear how a prediction previously validated to have a statistically significant correlation between an increase in platelet count ex vivo and in vivo is also a prediction of in vivo platelet production of the patient which incorporates the comparing of step f). Further, “an increase in platelet count ex vivo and in vivo is further indefinite because it is unclear what is being compared for the increase in platelet count. For example, it is unclear if Applicant intends to compare an increase in platelet count ex vivo to an increase in platelet count in vivo, such that the platelet counts for each of ex vivo and in vivo are increased between two undefined states/conditions, or whether applicant intends to compare an increase in platelet count ex vivo vs in vivo, such that the “increase” is referring to the ex vivo value compared to the in vivo value. If the former, then use of the relative term “increase” additionally renders the claim indefinite because the term “increase” is not defined by the claim, the specification does not provide a standard for ascertaining the requisite degree, and one of ordinary skill in the art would not be reasonably apprised of the scope of the invention.
Additionally, recitation of “expressed either as fold increase or absolute platelet numbers” in claims 1 and 12 is indefinite because it is unclear whether the prediction of in vivo platelet production is to be expressed either as fold increase or absolute platelet numbers, whether the validation is to be expressed either as fold increase or absolute platelet numbers, or whether the statistically significant correlation between an increase in platelet count ex vivo and in vivo is to be expressed either as fold increase or absolute platelet numbers. As above, use of the relative term “increase” additionally renders the claim indefinite because the term “increase” is not defined by the claim, the specification does not provide a standard for ascertaining the requisite degree, and one of ordinary skill in the art would not be reasonably apprised of the scope of the invention.
Therefore, the metes and bounds of the claims cannot be determined.
Independent claims 1 and 12 recite, “, with an R-squared value of at least 0.6 and a p value of less than 0.001 in a population study having at least 8 patients”, which is indefinite because it is unclear whether the prediction itself is with an R-squared value of at least 0.6 and a p value of less than 0.001 in a population study having at least 8 patients or whether the validation is with an R-squared value of at least 0.6 and a p value of less than 0.001 in a population study having at least 8 patients. As such, the metes and bounds of the claims cannot be determined.
Independent claim 1 recites the limitations “the patient” in lines 34 and 35. There is insufficient antecedent basis for this limitation in the claim. Independent claim 1 recites “a patient” in line 1 and “at least 8 patients” in lines 32-33. Therefore, it is unclear which patient “the patient” of lines 34 and 35 are referring to. As such, the metes and bounds of the claims cannot be determined. Claims 2-4 and 6-11 are included in this rejection due to their dependence on claims 1.
Claims 2 and 13 are indefinite in their recitation of “substantially free” since it is unclear how this term “substantially” is defined, what its metes and bounds are, or to what the term is directed towards. It is not clear in reference to what extent the two three-dimensional bone marrow models are free of tubular blood vessel mimicking structures. Therefore, the metes and bounds of the claims cannot be determined.
Claims 6 and 17 each recite, “wherein the patient has an ANKRD26-RT or MYH9-RD gene variant”, which is indefinite because it is unclear whether Applicant intends to claim wherein the patient has any gene variant associated with either of the diseases ANKRD26-RT or MYH9-RD, or whether Applicant intends to claim wherein the patient has gene variants specifically in the genes ANKRD26 or MYH9. Although Applicant recites the disease name, the name is italicized as though to indicate a gene name. As such, the metes and bounds of the claim cannot be determined.
Claims 8 and 19 each recite, “wherein the report includes megakaryocyte differentiation” in lines 1-2, which is indefinite because it is unclear how a report, which is a written collection of information, can include/comprise the differentiation of cells, which is a process undergone by cells. As such, the metes and bounds of the claim cannot be determined.
Claims 3-4, 7, 9-11, 14-16, 18, and 20-22 are included in this rejection due to their dependence on independent claim 1 and/or independent claim 12.
In the interest of compact prosecution, step g) has been interpreted such that the prediction incorporates both 1) the comparing of step f) and 2) a previously validated relationship between
a medicament-dependent increase in platelet counts generated ex vivo from patient-specific megakaryocytes compared to platelet counts generated ex vivo from control patient-specific megakaryocytes lacking the medicament and
a medicament-dependent increase in platelet counts generated in vivo within the patient administered the medicament compared to platelet counts generated in vivo within the patient having administered a control lacking the medicament,
wherein the patient-specific cells are cells specific for the patient receiving the medicament administration,
further wherein the relationship was validated in a population study having at least 8 patients, wherein the study produced results indicating a statistically significant correlation between a medicament-dependent increase in platelet count generated ex vivo produced by patient-specific megakaryocytes originating from the at least 8 patients compared to platelet counts generated ex vivo from control patient-specific megakaryocytes originating from the at least 8 patients lacking the medicament and a medicament-dependent increase in platelet count generated in vivo within the at least 8 patients compared to platelet counts generated in vivo within the at least 8 patients having received a control administration lacking the medicament,
wherein the correlation has an R-squared value of at least 0.6 and a p value of less than 0.001.
Claim Rejections - 35 USC § 112(a)
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 1-4 and 6-22 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, because the specification, while being enabling for
a) a patient-specific method of administering a medicament to a patient in need thereof, wherein the medicament is indicated to modulate megakaryocyte differentiation, proplatelet formation, and/or platelet production in the patient, and wherein the medicament is a thrombopoietin-receptor agonist; and
b) a patient-specific method of predicting in vivo efficacy of a thrombopoietin-receptor agonist as set forth in claim 12, wherein the medicament is a thrombopoietin-receptor agonist;
does not reasonably provide enablement for
a) a patient-specific method of administering a medicament to a patient in need thereof, wherein the medicament is indicated to modulate megakaryocyte differentiation, proplatelet formation, and/or platelet production in the patient, and wherein the medicament is any medicament; and
b) a patient-specific method of predicting in vivo efficacy of a thrombopoietin-receptor agonist as set forth in claim 12, wherein the medicament is any medicament. The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the invention commensurate in scope with these claims.
This rejection addresses the issue of an absence of an enabling disclosure for predicting in vivo efficacy of a thrombopoietin (TPO)-receptor agonist for a patient by introducing any medicament to the cells other than a TPO-receptor agonist. 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.
The specification does not provide an enabling disclosure for a) administering a medicament to a patient in need thereof, wherein the medicament is indicated to modulate megakaryocyte differentiation, proplatelet formation, and/or platelet production in the patient, and wherein the medicament is any medicament or b) predicting in vivo efficacy of a thrombopoietin-receptor agonist for a patient by introducing any medicament to the cells without adding a thrombopoietin-receptor agonist to the cells.
The broadest independent claim, claim 12, recite a patient-specific method of predicting in vivo efficacy of a thrombopoietin-receptor agonist for a patient, wherein the method comprises introducing any medicament to the cells in step c) and generating a report including a prediction of in vivo platelet production of the patient upon having the medicament administered. However, independent claim 12 does not recite any step directed to a thrombopoietin-receptor agonist. Claims 13-22 depend on claim 12.
Similarly, independent claim 1 recites a patient-specific method of administering a medicament to a patient in need thereof, wherein the medicament is indicated to modulate megakaryocyte differentiation, proplatelet formation, and/or platelet production in the patient, wherein the method comprises introducing any medicament to the cells in step c), generating a report including a prediction of in vivo platelet production of the patient upon having the medicament administered in step g), and then administering the medicament to the patient in step h). Claims 2-4 and 6-11 depend on claim 1.
The specification discloses predicting in vivo efficacy of a TPO-receptor agonist for a patient by introducing a TPO-receptor agonist (e.g., eltrombopag (EPAG)) ex vivo to cells obtained from the patient [0009, 0015, 0045, 0064, 0070, 0074-0076, Figure 5]. However, the specification does not disclose predicting in vivo efficacy of a TPO-receptor agonist for a patient by introducing any medicament other than a TPO-receptor agonist ex vivo to cells obtained from the patient.
The art at the time of filing teaches that TPO-receptor agonists function by stimulating platelet production through the TPO-receptor (c-Mpl) [column 1 ¶ 1- column 2 ¶ 1] and have distinct mechanisms of action, wherein even within the class of TPO-receptor agonists, eltrombopag and romiplostim have distinct mechanisms for increasing platelet output through distinct c-Mpl functions [Di Buduo et al. 2016, Haematologica, 101(12), 1479-1488, column 13 ¶ 1]. Specifically, eltrombopag increases platelet output by stimulating increased megakaryocyte maturation and proplatelet formation through upregulation of both pAKT and pERK, whereas romiplostim increases platelet output by increasing mainly megakaryocyte proliferation through upregulation of pAKT without a correspondent parallel stimulation of megakaryocyte maturation [column 13 ¶ 1- column 14 ¶ 1, Figure 7]. Therefore, even within the class of TPO-receptor agonists, treatment of one TPO-receptor agonist does not provide predictability of response to another TPO-receptor agonist. Accordingly, an ordinarily skilled artisan at the time of filing would not have expected a medicament which is not a TPO-receptor agonist, such as a medicament which does not act through the TPO-receptor and which is even less closely related mechanistically to a TPO-receptor agonist compared to other TPO-receptor agonists, to be able to predict the in vivo platelet production efficacy of a TPO-receptor agonist in a patient nor to modulate megakaryocyte differentiation, proplatelet formation and/or platelet production in the patient.
Neither the specification nor the art at the time of filing teaches predicting in vivo efficacy of a TPO-receptor agonist for a patient by introducing any medicament other than a TPO-receptor agonist ex vivo to cells obtained from the patient. Neither do the specification nor the art at the time of filing teach using that prediction of in vivo platelet production efficacy to administer any medicament other than a TPO-receptor agonist to the patient. Thus, in view of the specificity of TPO-receptor agonists to function through the TPO-receptor to stimulate responses in cells, the variety of TPO-receptor agonist mechanisms of action, and the breadth of the claims, the ordinarily skilled artisan would have considered predicting in vivo efficacy of a TPO-receptor agonist for a patient by introducing any medicament other than a TPO-receptor agonist ex vivo to cells obtained from the patient as highly unpredictable. Likewise, an ordinarily skilled artisan at the time of filing would have considered using the prediction so obtained to administer any medicament other than a TPO-receptor agonist to a patient as highly unpredictable. As such it would have required undue experimentation to practice the scope of applicant’s invention as claimed.
Claim Rejections - 35 USC § 103
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.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
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.
Claim(s) 1-4 and 6-22 are rejected under 35 U.S.C. 103 as being unpatentable over Di Buduo et al. [2016, Haematologica, 101(12), 1479-1488]; in view of Di Buduo et al. [2015, Blood, 125(14), 2254-2264, IDS]; Reems et al. [2010, Transfusion Medicine Reviews, 24(1), 33-43]; Silva et al. [2017, Cancer Research, 77(12), 3336-3351]; Majumder et al. [2015, Nature Communications, 6:6169, 1-14]; and Balduini et al. [2018, European Journal of Medical Genetics, 61(11), 715-722, IDS].
Note that the steps of a) preparing, b) seeding, c) introducing, d) perfusing, and e) maturing do not have any particular order indicated by claims 1 and 12 as written beyond that the cells are within the experimental model at the time of introducing the medicament. Specifically, the claims as written do not require that the introducing the medicament step is performed before the maturing step, nor do the claims require that the medicament be the agent for maturing the cells.
Regarding claims 1 and 12, Di Buduo (2016) teaches that eltrombopag modulates megakaryocyte differentiation, proplatelet formation, and platelet production ex vivo and at least modulates/increases platelet production in vivo [column 5 ¶ 1, column 11 ¶ 2].
Di Buduo (2016) also teaches a method comprising the following steps:
acquiring cells from a patient (e.g., human cord blood or peripheral blood samples) and preparing patient-specific hematopoietic stem cells (HSCs) therefrom [column 2 ¶ 2, column 7 ¶ 2];
seeding the HSCs into at least two 3-D bone marrow models,
wherein the at least two 3-D bone marrow models comprise a regenerated silk fibroin sponge having an interconnected network of pores with pore sizes and pore distributions adapted to facilitate cell adhesion, proplatelet extension, proplatelet formation, and platelet release and collection from maturing HSCs seeded therein [column 2 ¶ 2, column 7 ¶ 2, column 8 ¶ 2, Figure 3, 4],
wherein the at least two 3-D bone marrow models comprise an experimental model comprising eltrombopag and a control model lacking eltrombopag (e.g., comprising rHuTPO instead) [column 2 ¶ 2, column 7 ¶ 2, Figure 3], and
wherein each of the regenerated silk fibroin sponges comprise at least one protein of the extracellular matrix (e.g., the elected fibronectin) [Figure 4];
introducing the medicament (e.g., eltrombopag) to the HSCs within the experimental model [column 2 ¶ 2, column 7 ¶ 2, Figure 3];
perfusing the at least two 3-D bone marrow models with a cell culture medium [column 2 ¶ 2, column 7 ¶ 2, column 8 ¶ 2];
maturing the HSCs into megakaryocytes (e.g., HSCs were differentiated into megakaryocytes), wherein the megakaryocytes demonstrated cell adhesion, proplatelet extension, proplatelet formation, and platelet release, and wherein the platelets were collected [column 2 ¶ 2, column 7 ¶ 2- column 8 ¶ 2, Figure 3, 4]; and
comparing platelet production from the megakaryocytes within the experimental model versus megakaryocytes within the control model lacking eltrombopag [column 8 ¶ 2- column 9 ¶ 1, Figure 3, 4].
Di Buduo (2016) further teaches the administration of eltrombopag to subjects in need (e.g., having thrombocytopenia) for increasing platelet counts, including in subjects having inherited thrombocytopenia associated with mutations of the MYH9 gene [column 10 ¶ 2- column 11 ¶ 1]. Di Buduo (2016) also teaches that their study warrants further investigation on outcomes from the HSCs of patients affected by thrombocytopenias and overall bone marrow pathologies, such that understanding the efficacy and safety of thrombopoietin mimetics on pathologic samples before treating patients in vivo would represent an important step towards creating more personalized and effective therapies [column 13 ¶ 2], thereby teaching to use patient-specific cells in the method taught by Di Buduo (2016), wherein the patient is a patient in need of a medicament for increasing platelet production (e.g., eltrombopag). As such, Di Buduo (2016) teaches that the patient-specific cells originate from a patient in need of eltrombopag.
Di Buduo (2016) does not explicitly teach all of the limitations of steps g) and h) nor the elected species of patient-specific megakaryocyte progenitors.
Regarding steps g) and h), Di Buduo (2016) teaches that understanding the efficacy and safety of thrombopoietin mimetics (e.g., eltrombopag) on pathologic samples before treating patients in vivo would represent an important step towards creating more personalized and effective therapies [column 15 ¶ 2]. Di Buduo (2016) also teaches to present platelet production as an absolute platelet number [Figure 4].
Di Buduo (2015) teaches comparing ex vivo platelet production from patient-specific megakaryocytes with in vivo platelet production in the same patients from which the patient-specific megakaryocytes are derived, wherein the results were consistent with their previous finding demonstrating a significant correlation between in vivo platelet count and in vitro proplatelet formation from patients with myeloproliferative neoplasms [column 13 ¶ 1- column 14 ¶ 1, Figure 8]. Di Buduo (2015) also teaches a significant correlation among proplatelet branching along the silk vascular tube, the extent of platelet release ex vivo, and in vivo platelet count was observed when MKs derived from patients with primary myelofibrosis were seeded in the bone marrow model [column 17 ¶ 2- column 18 ¶ 1]. Di Buduo (2015) also teaches that patient-derived MKs and endothelial cells can be cultured in the silk sponge 3D bone marrow model and used to design patient-specific drug administration regimes [column 18 ¶ 2].
Therefore, given the teaching of Di Buduo (2016) that understanding the efficacy of eltrombopag on pathologic samples before treating patients in vivo is important for creating more personalized and effective therapies; and the teachings of Di Buduo (2015) to compare ex vivo platelet production from patient-specific megakaryocytes with in vivo platelet production in the same patients, of a significant correlation between the extent of platelet release ex vivo and in vivo platelet count for MKs derived from patients, and to design patient-specific drug administration regimes therefrom; an ordinarily skilled artisan would have been motivated to use the correlation between ex vivo platelet release and in vivo platelet counts in a method to design patient-specific drug administration regimes, wherein a patient-specific ex vivo response to eltrombopag is predictive of a patient-specific in vivo response to eltrombopag, for more personalized and effective therapies.
The combined teachings of Di Buduo (2016) and Du Buduo (2015) do not teach wherein the prediction is validated with an R-squared value of at least 0.6 and a p value of less than 0.001 in a population study having at least 8 patients.
However, Silva teaches an ex vivo platform for the prediction of clinical responses in multiple myeloma [title]. Silva teaches that their system consists of patient-specific mathematical models parameterized by an ex vivo assay that allows them to predict clinical response to up to 31 drugs within 5 days after bone marrow biopsy [abstract]. Silva further teaches that they observed a significant correlation between predicted and actual tumor burden measurements with a p-value < 0.0001 [abstract].
Silva further teaches generating a report, wherein the report includes a prediction of the in vivo response of a patient upon having a medicament/drug administered [column 6 ¶ 2, Supplemental Data Processing Guide page 1, 6, 7]. Silva teaches that the prediction incorporates the comparing of the drug response resulting from treatment of patient-specific cells ex vivo to a response resulting from treatment of patient-specific cells with a control lacking the drug [column 5 ¶ 4]. Silva also teaches that the predication is validated in a population study having 52 patients (e.g., at least 8 patients) [abstract, column 8 ¶ 2-3], wherein the study produced results indicating a statistically significant correlation between a drug-dependent response generated ex vivo in patient-specific cells originating from the 52 patients and a drug-dependent response generated in vivo within the 52 patients [column 5 ¶ 4, column 6 ¶ 2-7, column 7 ¶ 2- column 8 ¶ 3, Table 1]. Silva also teaches that the correlation has a p value of less than 0.001 [abstract, column 11 ¶ 2, Figure 3].
Silva further teaches that their approach can provide precise clinical insight about treatment efficacy in a timely manner and assist physicians in practicing truly personalized management by proposing the best choice of therapy for each patient [column 17 ¶ 2]. Silva further teaches using the prediction as a decision-support system for choice of therapy to identify the right drug for the right patient at the right time, and that their approach may be used to assess patient sensitivity to targeted therapeutics facilitating patient sample-derived drug screening [column 11 ¶ 3- column 12 ¶ 2, Figure 4].
Therefore, given the teachings of Silva to generate a report which includes a prediction validated in a population study having greater than 8 patients, wherein the study produced results indicating a statistically significant correlation with a p-value less than 0.001 between a drug-dependent response generated ex vivo in patient-specific cells and a drug-dependent response generated in vivo; and the teaching of Silva that their approach can assist physicians in practicing truly personalized management by proposing the best choice of therapy for each patient; an ordinarily skilled artisan would have been motivated to generate a report comprising a prediction of the in vivo drug-dependent response of a patient which incorporates the comparing of an ex vivo patient-specific drug-dependent response with an ex vivo patient-specific control sample lacking the drug, and wherein the prediction has been validated to have a statistically significant correlation between an ex vivo drug-dependent response and an in vivo drug-dependent response, wherein the statistical significance is demonstrated by a population study having at least 8 patients and producing a p-value indicative a statistical significance (e.g., p < 0.001), to facilitate personalized treatment. Given Silva’s teachings to use the information from the test as a decision-support system for choice of therapy, an ordinarily skilled artisan at the time of filing would have been additionally motivated to use the results of the test to determine administration of a medicament to the patient.
The combined teachings of Di Buduo (2016) and Du Buduo (2015) and Siva do not teach wherein the prediction is validated with an R-squared value of at least 0.6 in the population study having at least 8 patients nor administering the medicament to the patient if the prediction exceeds a predetermined threshold and forgoing administering the medicament to the patient if the prediction fails to exceed the predetermined threshold.
However, Majumder teaches predicting clinical response to anticancer drugs using an ex vivo platform [title]. Majumder further teaches validating their method (e.g., CANScript) for predicting response to cytotoxic and targeted drugs by comparing drug response in human tumor-derived xenotransplants (HTX) (i.e., in vivo drug response in the patient) and in matched CANScripts explants derived from the HTX (i.e., ex vivo drug response of cells obtained from the patient), resulting in excellent correlation between the in vivo and ex vivo outcomes, wherein the linear correlation between in vivo and ex vivo responses from at least 10 patients has an R-squared value of 0.903 indicative of an excellent correlation [column 11 ¶ 2- column 15 ¶ 1, Figure 6]. Majumder further teaches that the robustness of their platform in predicting clinical response could potentially be useful for personalizing treatment [column 1 ¶ 3], and that the CANScript method can afford nearly high-throughput testing, allowing predictions to be made within 7 days for truly personalizing chemotherapy [column 18 ¶ 2].
Majumder further teaches determining a threshold value for the prediction, wherein a threshold value of >19.1 in their system is indicative of a predicted partial responder and a threshold value of >55.14 is indicative of a predicted complete responder [column 15 ¶ 2-3]. Majumder further teaches that combining functional read-outs together with predetermined thresholds resulted in predictions that were significantly better than what could have been achieved by predicting using any one of the functional read-outs alone [column 15 ¶ 2]. Majumder also teaches that the prediction tool can emerge as a powerful platform for enabling personalized medicine [abstract].
Therefore, given the teachings of Majumder for predicting clinical response to anticancer drugs using an ex vivo platform such that functional validation of the prediction resulted in excellent correlation between the outcomes in the ex vivo platform and the response to chemotherapy in the in vivo studies, wherein the linear correlation between ex vivo outcomes and in vivo responses from at least 10 patients has an R-squared value of 0.903; and the further teachings of Majumder that the robustness of their platform in predicting clinical response could be useful for truly personalizing chemotherapy; an ordinarily skilled artisan would have been motivated to use a prediction which has been validated to have an R-squared value indicative of an excellent correlation (e.g., R2 = 0.903), to provide precise clinical insight about treatment efficacy in a timely manner to facilitate personalized treatment. Additionally, an ordinarily skilled artisan would have been further motivated to determine a threshold value for classification of drug response to determine whether or not a patient’s condition will respond favorably to a drug, and therefore to use the predicted response in deciding whether or not to administer the drug to the patient, thereby identifying the right drug for the right patient.
Regarding the elected species of megakaryocyte progenitors, Di Buduo (2016) also teaches that megakaryocytes efficiently differentiated from their progenitors when concentrations of eltrombopag in the same range as those measured in the serum of healthy subjects during oral eltrombopag administration were used in culture [column 5 ¶ 1].
Additionally, Di Buduo (2015) teaches the seeding of mature megakaryocytes derived from peripheral blood progenitors, wherein MKs successfully generated platelets ex vivo and similar platelet production was obtained using hematopoietic progenitor cells from different sources, including patient-derived MKs [column 4 ¶ 1-2, 4, column 8 ¶ 1, column 12 ¶ 3- column 14 ¶ 1, column 15 ¶ 1, column 17 ¶ 2, supplemental methods page 4 ¶ 3, supplemental methods page 5 ¶ 1, Figure 3, supplemental figure 2].
Further, Reems teaches that the differentiation of hematopoietic stem cells (HSCs) into megakaryocytes involves the differentiation of HSCs into megakaryocyte progenitors, which them differentiate into megakaryocytes. Accordingly, by teaching the differentiation of hematopoietic stem cells (HSCs) into megakaryocytes, Di Buduo (2016) is teaching the differentiation of patient-specific HSCs into patient-specific megakaryocyte progenitors and then into patient-specific megakaryocytes, thereby teaching preparing patient-specific megakaryocyte progenitors from cells obtained from the patient and maturing the patient-specific megakaryocyte progenitors into patient-specific megakaryocytes.
Therefore, given the teachings of Di Buduo (2016) that megakaryocytes efficiently differentiated from their progenitors in the presence of the TPO mimetic eltrombopag; the teachings of Di Buduo (2015) to seed mature megakaryocytes into the silk sponges after in vitro differentiation from HSCs; and the teachings of Reems that megakaryocyte progenitors are an intermediate cell state in the differentiation of HSCs into megakaryocytes, it would have been obvious to an ordinarily skilled artisan at the time of filing the instant application to substitute any of the cells along the differentiation pathway from HSCs to mature megakaryocytes, including megakaryocyte progenitors, for the HSCs taught by Di Buduo with a reasonable expectation of success in obtaining eltrombopag-induced platelet production therefrom, wherein selection of megakaryocyte progenitors as the cells to seed amounts to selecting from a limited number of options of cells which are taught to mature to megakaryocytes.
Regarding claims 2 and 13, Di Buduo (2016), Di Buduo (2015), Reems, Silva, and Majumder teach the limitations of independent claims 1 and 12. Di Buduo (2016) does not teach that the silk fibroin 3D bone marrow models comprise any tubular vessel mimicking structures. Therefore, absent evidence to the contrary, the 3D bone marrow models of Di Buduo (2016) are considered to be substantially free of tubular blood vessel mimicking structures.
Regarding claims 3-4 and 14-16, Di Buduo (2016), Di Buduo (2015), Reems, Silva, and Majumder teach the limitations of independent claims 1 and 12. As discussed above, Di Buduo (2016) teaches that the medicament is a thrombopoietin-receptor agonist, eltrombopag.
Regarding claims 6 and 17, Di Buduo (2016), Di Buduo (2015), Reems, Silva, and Majumder teach the limitations of independent claims 1 and 12. Additionally, Balduini teaches that mutations in the ANKRD26 gene cause ANKRD26-related thrombocytopenia (i.e., ANKRD26-RT), which is an inherited thrombocytopenia, wherein the thrombocytopenia is a consequence of a defect in megakaryocyte differentiation and/or maturation [column 3 ¶ 2-3]. Therefore, a patient having ANKRD26-RT is a patient in need of a medicament indicated to modulate megakaryocyte differentiation, proplatelet formation, and/or platelet production. Balduini also teaches that eltrombopag is a TPO nonpeptide mimetic which has been approved for treating ITP and for increasing platelet count in thrombocytopenia due to hepatitis C infection [column 9 ¶ 3].
Therefore, an ordinarily skilled artisan at the time of filing the instant application would have been motivated to use the patient-specific method of administering a medicament to a patient in need thereof, wherein the medicament is indicated to modulate megakaryocyte differentiation, proplatelet formation, and/or platelet production, for a patient having ANKRD-RT to counter the patient’s defect in megakaryocyte differentiation and/or maturation, and in so doing would predict the in vivo efficacy of a TPO-receptor agonist, such as eltrombopag, for the patient.
Regarding claims 7 and 18, Di Buduo (2016), Di Buduo (2015), Reems, Silva, and Majumder teach the limitations of independent claims 1 and 12. Di Buduo (2016) also teaches the analysis of megakaryocyte differentiation of the patient-specific megakaryocytes [column 3 ¶ 2, column 4 ¶ 1-column 6 ¶ 1, Figure 1].
Regarding claims 8 and 19, Di Buduo (2016), Di Buduo (2015), Reems, Silva, and Majumder teach the limitations of independent claims 1 and 12. As discussed above, Di Buduo (2016) teaches the analysis of megakaryocyte differentiation of the patient-specific megakaryocytes in the presence of the TPO-receptor agonist eltrombopag [Figure 1]. Balduini teaches that mutations in the ANKRD26 gene cause ANKRD26-related thrombocytopenia (i.e., ANKRD26-RT), which is an inherited thrombocytopenia, wherein the thrombocytopenia is a consequence of a defect in megakaryocyte differentiation and/or maturation [column 3 ¶ 2-3]. Therefore, an ordinarily skilled artisan at the time of filing the instant application would have been motivated to include information regarding megakaryocyte differentiation and/or maturation in a report on the effects of eltrombopag on patient-specific megakaryocyte progenitor cells, along with the prediction of in vivo platelet production, for a patient having ANKRD-RT to assess the effect of eltrombopag treatment on countering the patient’s defect in megakaryocyte differentiation and/or maturation.
Regarding claims 9 a