METHODS FOR ASSESSING GRAFT SUITABILITY FOR TRANSPLANTATION
This invention relates to methods and compositions for assessing the suitability of a graft for transplantation by measuring total and/or specific cell-free nucleic acids (such as cf-DNA) and/or cell lysis. Specifically, the method comprising obtaining an amount of total short fragment cf-DNA and/or graft-specific short fragment cf-DNA released from a potential graft (e.g., ex vivo), e.g., prior to contacting of the potential graft with blood cells of a potential recipient, and/or subsequent to contacting of the potential graft or cells thereof with blood cells from a potential recipient, and assessing the amount(s) to determine the suitability of the potential graft
1 . A method of assessing the suitability of a potential graft for transplantation, comprising:
a. obtaining an amount of total short fragment cf-DNA and/or graft-specific short-fragment cf-DNA released from a graft prior to contacting the graft with blood cells from a potential recipient, and
b. assessing the amount(s) to determine the suitability of the graft for transplantation.
2 . The method of claim 1 , wherein obtaining the amount comprises performing a method to quantify the amount of total short fragment cf-DNA and/or graft-specific short fragment cf-DNA released by the graft.
3 . The method of claim 1 wherein the method further comprises performing a method to quantify the amount of total short fragment cf-DNA and/or graft-specific short-fragment cf-DNA released by the graft subsequent to contacting the graft or cells thereof with blood cells from a potential recipient.
4 . A method of assessing the suitability of a graft for transplantation, comprising:
a. obtaining an amount of total short fragment cf-DNA and/or graft-specific short fragment cf-DNA released by the graft subsequent to contacting of the graft or cells thereof with blood cells from a potential recipient, and
b. assessing the amount(s) to determine the suitability of the potential graft for transplantation.
5 . The method of claim 4 , wherein obtaining the amount comprises performing a method to quantify the amount(s) of total short fragment cell-free DNA and/or graft-specific short fragment cf-DNA in order to obtain the amount.
6 . The method of claim 1 , wherein the method further comprises determining an amount of cell lysis.
7 . The method of claim 6 , wherein the determining the amount of cell lysis comprises performing fragment analysis, wherein the fragment analysis comprising quantifying long and/or short fragments.
8 . (canceled)
9 . The method of claim 1 , wherein the method further comprises one, two, three or more centrifugation steps of the sample(s) prior to obtaining the amount of total short fragment cf-DNA and/or graft-specific short-fragment cf-DNA and/or cell lysis.
10 . (canceled)
11 . The method of claim 1 , wherein the method further comprises obtaining an amount of total short fragment cf-DNA and/or graft-specific short-fragment cf-DNA and/or determining the amount of cell lysis at one or more additional time points.
12 . The method of claim 1 , wherein the method further comprises comparing the amount(s) of the total short fragment cf-DNA and/or graft-specific cf-DNA and/or amount of cell lysis with threshold value(s) or amount(s) obtained from additional time point(s).
13 . The method of claim 1 , wherein the amount(s) of the total short fragment cf-DNA and/or graft-specific cf-DNA and/or amount of cell lysis are obtained in a sample of media in which the potential graft or cells therefrom are contained or are in contact with.
14 . The method of claim 13 , wherein the media is a storage, perfusion, or preservation media in which the graft is contained or in contact with, wherein the cf-DNA is released from the graft into the media.
15 . (canceled)
16 . The method of claim 1 , wherein the amount(s) is/are obtained in a perfusate sample or an EVLP sample.
17 .- 18 . (canceled)
19 . The method of claim 1 , wherein the potential graft or cells thereof are from a potential donor of the same species as the potential recipient.
20 . The method of claim 1 , wherein the potential graft is an organ or organs comprising a heart, a lung, or a heart and a lung.
21 - 23 . (canceled)
24 . The method of claim 1 , wherein the graft is monitored every 15 minutes, every 30 minutes, hourly, daily, weekly, bimonthly or monthly prior to transplantation.
25 . (canceled)
26 . The method of claim 1 , wherein the total short fragment cell-free DNA and/or graft-specific short fragment cf-DNA and/or cell lysis is measured using a quantitative amplification method.
27 . (canceled)
28 . The method of claim 1 , wherein the amount of total short fragment cell-free DNA and/or graft-specific short fragment cf-DNA and/or cell lysis is measured using next-generation sequencing.
29 . The method of claim 1 , wherein when the value for the amount(s) are above a threshold value or a value from a prior point in time, decreasing suitability of the graft or unsuitability of the graft for transplantation is indicated; or wherein when the value for the amount(s) are below a threshold value or a value from a prior point in time, increasing suitability of the graft or suitability of the graft.
30 . (canceled)
31 . The method of claim 1 , further comprising performing treatment of the graft, potential donor, and/or recipient.
32 - 33 . (canceled)