Pig Kidney Xenotransplantation Research Articles

315.2: Considerations for inclusion/exclusion criteria for clinical pig kidney xenotransplantation

315.2: Considerations for inclusion/exclusion criteria for clinical pig kidney xenotransplantation

Assessment of glomerular filtration and tubular secretion in baboons with life-supporting pig kidney grafts.

With pig kidney xenotransplantation nearing clinical reality, it is imperative to measure pig kidney function in the graft recipients. Our aims were (i) to compare inulin clearance after a short intravenous (IV) bolus with steady-state inulin IV infusion, (ii) to use this method to measure the glomerular filtration rate (GFR), and (iii) to determine the tubular secretory function using cefoxitin in a pig-to-baboon renal transplant model. A short IV infusion of inulin and cefoxitin were followed by a maintenance IV infusion of inulin over 5 h in seven healthy baboons, three healthy pigs, and five baboons after bilateral native nephrectomy and intra-abdominal pig renal transplantation. Blood and urine samples were collected. Serum and urinary inulin and serum cefoxitin concentrations measured by validated assays were used to calculate GFR and renal secretion. GFR calculatedwere similar by both methods. The body weight normalized total body clearance of inulin was similar in pigs and baboons despite differences in absolute clearances. Pig kidney transplanted into baboons provided similar clearance in baboons when normalized to baboon body weight and sustained filtration and secretory functions. The study documented that pig kidneys support the physiologic needs of baboons and are likely to support human recipients as well.

Renin-angiotensin-aldosterone system function in the pig-to-baboon kidney xenotransplantation model

After pig-to-baboon kidney transplantation, episodes of hypovolemia and hypotension from an unexplained mechanism have been reported. This study evaluated the renin-angiotensin-aldosterone system post–kidney xenotransplantation. Kidneys from genetically-engineered pigs were transplanted into 5 immunosuppressed baboons after the excision of the native kidneys. Immunosuppressive therapy was based on the blockade of the CD40/CD154 costimulation pathway. Plasma renin, angiotensinogen (AGT), angiotensin II (Ang II), aldosterone levels, and urine osmolality and electrolytes were measured in healthy pigs, healthy nonimmunosuppressed baboons, and immunosuppressed baboons with life-supporting pig kidney grafts. After pig kidney transplantation, plasma renin and Ang II levels were not significantly different, although Ang II trended lower, even though plasma AGT and potassium were increased. Plasma aldosterone levels were unchanged. Urine osmolality and sodium concentration were decreased. Even in the presence of increasing AGT and potassium levels, lower plasma Ang II concentrations may be because of reduced, albeit not absent, the reactivity of pig renin to cleave baboon AGT, suggesting an impaired response of the renin-angiotensin-aldosterone system to hypovolemic and hypotensive episodes. The maintenance of aldosterone may be protective. The reduced urine osmolality and sodium concentration reflect the decreased ability of the pig kidney to concentrate urine. These considerations should not prohibit successful clinical pig kidney xenotransplantation.

The Polemic of Pig Kidney Xenotransplantation for Human Life Sustainability:

End-stage renal disease (ESRD) cases continue to increase. However, the increase in cases of ESRD is not matched by the supply of kidneys as a condition for being able to perform a kidney transplant. Although this is not the only way to treat this case, it is only with a kidney transplant that the patient can truly recover. New York University Langone's success in xenotransplantation procedure using a genetically engineered pig kidney has become a new polemic in Islamic law issues. According to Islamic law, a pig has forbidden legal status (haram). This article analyzes the practice of xenotransplantation of pig kidneys in the human body using the concept of “Halal-haram” by Yusuf al-Qaradawi and “Maslahah” by Harun Sencal & Mehmet Asutay. This study is a qualitative method using a library research approach. This study found that it is permissible to do prohibited things during an emergency (daruriyat). In addition, in the realm of maslahah, hajiyat cannot beat daruriyat. Finally, summing up from these two Islamic scholars’ thoughts indicates that pig kidney xenotransplantation is prohibited as long as other alternatives to treat ERSD patients are available. Some options might be taken, such as hemodialysis and blood filtration (CAPD); therefore, it cannot be considered an emergency.
 Keywords: Pig kidney xenotransplantation, Islamic legal thoughts, halal-haram concept, principle of mashlahah   

Pig Kidney Xenotransplantation as an Alternative Solution of Hifdz Al Nafs

Recently, US surgeons successfully tested pig kidney transplants in a human patient, which caught the medical community by surprise. This has elicited a range of responses, including among Muslim communities, whether it is permitted by shariah or vice versa. Some Muslim scholars stated that pigs are prohibited animals under Islamic law. In contrast, others saw this success as the most recent alternative treatment for kidney failure in recent history. This research examines the issue of transplanting pig kidneys into human bodies by maqasid al-syari'ah (hifdz al-nafs). The full qualitative method was employed, which was based on library research to investigate the stated problems, and all the findings were analyzed descriptively. The study shows, first and foremost, a pig is one kind of animal that is prohibited under Islamic law in any form. Two conditions must be fulfilled before the transplant of a pig's kidney into a human body can be justified: (1) it must be performed in an emergency where there is no alternative medicine or other sacred organs available; and (2) the harm resulting from the transplant itself must be less than the harm resulting from not performing the transplant. Thus, the transplantation of a pig's kidney into a human body is part of the hifdz al nafs effort to implement maqasid al dharuriyyat for the patient's survival.

Pig-to-human kidney transplantation using brain-dead donors as recipients: One giant leap, or only one small step for transplantkind?

Pig kidney xenotransplantation is increasingly regarded as a realistic solution to the current shortage of human organ donors for patients with end-stage organ failure. Recently, the news of three pig-to-human transplantation cases has awakened public interest. Notably, the case by the Alabama team reported detailed and important findings for the xenotransplantation field. Using a genetically modified pig, two porcine kidneys were transplanted into a brain-dead recipient. They applied several approaches established in the preclinical NHP study, including gene-edited pig kidney graft and preoperative laboratory inspection such as crossmatching and infection screening. The pig-to-human kidney xenotransplantation had no unexpected events during surgery or evidence of hyperacute rejection. Unfortunately, the grafts did not work appropriately, and the study had to be terminated due to the decompensation of the recipient. While this study demonstrated the outstanding achievement in this research area, it also revealed remaining gaps to move xenotransplantation to the clinic. While brain-dead human recipients could reinforce the compatibility achievements of gene-edited pigs in NHP, their pro-inflammatory and pro-coagulant environment, in combination with short-duration of experiments will limit the assessment of kidney function, infection and rejection risk post-transplant, in particular antibody-mediated rejection. The use of successful immunosuppressive protocols of non-human primates xenotransplant experiments including anti-CD154 antibody will be critical to maximize the success in the first in-human trials.

Informed Consent for Potential Recipients of Pig Kidney Xenotransplantation in the United States.

Clinical trials of kidney xenotransplantation are being considered in the United States. Before this novel procedure can take place, investigators will have to obtain approval from the institutional review board. The consent form that will be used for such a trial and that will receive approval from the institutional review board will be complex. Informed consent-the process by which a research participant provides his/her permission to participate in a clinical trial-is a staple of the research process and most commonly is in the form of a physical document. In the case of a novel procedure with uncertain benefits and risks and a participant population in acute need of a transplant, the consent process is crucial. These complexities may raise several ethical considerations for the initial pig kidney xenotransplantation recipients in the United States that will require adaptations of the required elements of the informed consent process by the US Department of Human and Health Services. The ethical issues include (1) a subject's ability to withdraw from the trial, (2) restrictions on their reproductive rights, and (3) the possibility of the need for quarantine if there is a perceived risk of xenozoonosis. This article aims to discuss ethical considerations that may challenge the general required elements of the informed consent form stipulated by the 45 Code of Federal Regulations 46 of the US Department of Health and Human Services and to suggest recommendations for deliberation.

Both Natural and Induced Anti-Sda Antibodies Play Important Roles in GTKO Pig-to-Rhesus Monkey Xenotransplantation.

Sda, produced by the B4GALNT2 enzyme, has been recognized as an important xenoantigen for pig-to-nonhuman primate xenotransplantation. However, little is known about Sda expression in pigs and its immunogenicity in xenotransplantation. In this study, peripheral blood mononuclear cells (PBMCs) were isolated from wildtype, GTKO (with high, moderate, and low Sda expression), GTKO/β4GalNT2KO, GTKO/CMAHKO, or GTKO/CMAHKO/β4GalNT2KO pigs. Anti-pig IgM/IgG binding and complement-dependent cytotoxicity (CDC) to pig PBMCs was measured by flow cytometry using pooled rhesus monkey sera (n=20) or human sera (n=20). As compared to wild-type pigs (n=12), GTKO pigs (n=17) had a significantly higher mean level of Sda expression on PBMCs and showed a greater individual difference in expression. Both the overall binding of monkey serum IgM/IgG antibody to GTKO pig PBMCs and CDC against these PBMCs decreased significantly with a progressive reduction in Sda expression, showing a clear dose-effect relationship. Both the monkey serum antibody binding and CDC decreased significantly after the additional deletion of Sda, whereas the binding of human serum antibody and CDC against the GTKO pig PBMCs were markedly reduced after the deletion of Neu5Gc in the pigs. In addition, anti-Sda antibody accounted for > 50% of the induced anti-non-Gal antibody at the time of rejection in two rhesus monkeys that received GTKO/hCD55 pig kidney xenotransplantation, and the anti-Sda antibody showed significant cytotoxic activity against GTKO pig cells. We conclude that both natural and induced anti-Sda antibodies play important roles in GTKO pig-to-rhesus monkey xenotransplantation, thus providing further evidence for GTKO/β4GalNT2KO pigs as the preferred organ source for rhesus monkeys as a preclinical model of xenotransplantation.

Physiological aspects of pig kidney xenotransplantation and implications for management following transplant.

Successful organ transplantation between species is now possible, using genetic modifications. This article aims to provide a comprehensive overview of the differences and similarities in kidney function between humans, primates, and pigs, in preparation for pig-allograft to human xenotransplantation. The kidney, as the principal defender of body homeostasis, acts as a sensor, effector, and regulator of physiologic feedback systems. Considerations are made for anticipated effects on each system when a pig kidney is placed into a human recipient. Discussion topics include anatomy, global kidney function, sodium and water handling, kidney hormone production and response to circulating hormones, acid-base balance, and calcium and phosphorus handling. Based on available data, pig kidneys are anticipated to be compatible with human physiology, despite a few barriers.

Serum Antibody Binding and Cytotoxicity to Pig Cells in Chinese Subjects: Relevance to Clinical Renal Xenotransplantation.

Kidney xenotransplantation is expected to contribute to resolving the shortage of kidneys from deceased human donors. Although progress in experimental life-supporting pig renal xenotransplantation has been encouraging, there are still issues to be considered before a clinical trial can be initiated. We attempted to clarify some of these by an in vitro study. Blood was drawn from healthy volunteers (Volunteers, n=20), patients with end-stage renal disease (ESRD, n=20) pre-operation (Pre), and on Day 1 (POD 1) and Day 14 (POD 14) after renal allotransplantation, brain-dead organ donors (DBD, n=20), and renal allotransplant recipients who were currently experiencing T cell-mediated rejection (Allo-TCMR, n=20). Serum IgM/IgG binding to, and complement-dependent cytotoxicity (CDC) of, PBMCs and RBCs from (a) wild-type (WT), (b) α1,3-galactosyltransferase gene-knockout (GTKO), (c) GTKO/beta-1,4-N-acety1 galactosaminyltransferase 2-knockout (GTKO/β4GalNT2KO), (d) GTKO/cytidine monophosphate-N-acetylneuraminic acid hydroxylase-knockout (GTKO/CMAHKO), and (e) GTKO/β4GalNT2KO/CMAHKO/hCD55 (TKO/hCD55) pigs were measured by flow cytometry. We obtained the following results: (i) Serum IgM/IgG binding and CDC in Volunteers were significantly greater to WT, GTKO, and GTKO/β4GalNT2KO PBMCs or RBCs than to GTKO/CMAHKO and TKO/hCD55 cells; (ii) ESRD, DBD, and Allo-TCMR serum antibody binding and CDC to WT pig PBMCs were significantly greater than to GTKO, GTKO/β4GalNT2KO, GTKO/CMAHKO, and TKO/hCD55 cells; (iii) antibody binding to GTKO/CMAHKO pig cells was significantly lower in hemodialysis than peritoneal dialysis patients. (iv) Two of twenty allotransplantation recipients’ serum IgG binding to GTKO pig PBMCs increased on POD14 compared with Pre, but IgG binding to GTKO pig RBCs did not; (v) In all sera, the lowest antibody binding and CDC were to GTKO/CMAHKO and TKO/CD55 pig cells. We conclude (i) CMAHKO in the pig may be critical to the success of clinical pig kidney xenotransplantation, and may be the most important after GTKO, at least in Chinese patients; (ii) subjects with ESRD, or who are immunosuppressed after kidney allotransplantation, and DBD, have lower levels of antibody binding and CDC to genetically-engineered pig cells than do volunteers; (iii) TKO pigs with selected human ‘protective’ transgenes, e.g., CD55, are likely to prove to be the optimal sources of kidneys for clinical xenotransplantation.

Recent progress in the pig-to-nonhuman primate kidney transplantation model: Report of a symposium.

Three excellent presentations at an industry-sponsored symposium at the (virtual) congress of the combined IXA/CTRMS (September 23-25, 2021) were directed to the value and limitations of the pig-to-nonhuman primate (NHP) kidney transplantation model. Daniel Firl and James Markmann provided a meta-analysis comparing the results of kidney allotransplantation and xenotransplantation in NHPs during the past 25 years. Remarkably, the authors had identified 73 published reports that included 910 individual experiments. Although recipient survival after allotransplantation was longer, the superiority over the survival of xenografts was less than anticipated. Given the excellent short- and medium-term results of clinical kidney allotransplantation today, these data provide hope that the results of clinical pig kidney xenotransplantation may prove significantly better than in NHPs. The authors identified several factors that were shown to statistically influence the success or failure of xenotransplantation. Jean Kwun provided valuable information relating to the longstanding question of whether the survival of a pig organ would be jeopardized if transplanted into an allosensitized recipient. He demonstrated that pig kidney transplantation in an HLA-sensitized patient may be at a disadvantage, although multiple genetic engineering of the organ-source pig significantly delayed rejection. In the initial clinical trials, therefore, it would seem wise to exclude any patient with evidence of anti-HLA antibodies. Andrew Adams reported longer survival (>1 year) of Rhesus monkeys with life-supporting pig kidney grafts than has been achieved previously. Although not consistently achieved, these excellent results were obtained with an anti-CD154mAb-based regimen after CD4+ T cell and partial CD20+ B cell depletion. Factors that might have contributed to this success, including the phenotype of the pig, the species of the recipient, the recipient's anti-pig antibody level, and the immunosuppressive regimen, were discussed. Importantly, pig kidney function appeared to be normal in long-term surviving monkeys. Each study contributed to our goal of introducing xenotransplantation into the clinic.

"You cannot stay in the laboratory forever"*: Taking pig kidney xenotransplantation from the laboratory to the clinic.

Progress in life-supporting kidney transplantation in the genetically-engineered pig-to-nonhuman primate model has been encouraging, with pig kidneys sometimes supporting life for > 1 year. What steps need to be taken by (i) the laboratory team, and (ii) the clinical team to prepare for the first clinical trial? The major topics include (i) what currently-available genetic modifications are optimal to reduce the possibility of graft rejection, (ii) what immunosuppressive therapeutic regimen is optimal, and (iii) what steps need to be taken to minimize the risk of transfer of an infectious microorganism with the graft. We suggest that patients who are unlikely to live long enough to receive a kidney from a deceased human donor would benefit from the opportunity of a period of dialysis-free support by a pig kidney, and the experience gained would enable xenotransplantation to progress much more rapidly than if we remain in the laboratory.

Suggested Patient Selection Criteria for Initial Clinical Trials of Pig Kidney Xenotransplantation in the United States.

Suggested Patient Selection Criteria for Initial Clinical Trials of Pig Kidney Xenotransplantation in the United States.

Immunological selection and monitoring of patients undergoing pig kidney transplantation.

Pig kidney xenotransplantation has the potential to alleviate the current shortage of deceased and living human organs and provide patients with end-stage renal disease with a greater opportunity for long-term survival and a better quality of life. In recent decades, advances in the genetic engineering of pigs and in immunosuppressive therapy have permitted the resolution of many historical obstacles to the success of pig kidney transplantation in nonhuman primates. Pig kidney xenotransplantation may soon be translated to the clinic. Given the potential risks of kidney xenotransplantation, particularly of immunologic rejection of the graft, potential patients must be carefully screened for inclusion in the initial clinical trials and immunologically monitored diligently post-transplantation. We provide an overview of the immunological methods we believe should be used to (i) screen potential patients for the first clinical trials to exclude those with a higher risk of rejection, and (ii) monitor patients with a pig kidney graft to determine their immunological response to the graft.

Pig kidney xenotransplantation: Progress toward clinical trials.

Pig organ xenotransplantation offers a solution to the shortage of deceased human organs for transplantation. The pathobiological response to a pig xenograft is complex, involving antibody, complement, coagulation, inflammatory, and cellular responses. To overcome these barriers, genetic manipulation of the organ-source pigs has largely been directed to two major aims-(a) deletion of expression of the known carbohydrate xenoantigens against which humans have natural (preformed) antibodies, and (b) transgenic expression of human protective proteins, for example, complement- and coagulation-regulatory proteins. Conventional (FDA-approved) immunosuppressive therapy is unsuccessful in preventing an adaptive immune response to pig cells, but blockade of the CD40:CD154 costimulation pathway is successful. Survival of genetically engineered pig kidneys in immunosuppressed nonhuman primates can now be measured in months. Non-immunological aspects, for example, pig renal function, a hypovolemia syndrome, and rapid growth of the pig kidney after transplantation, are briefly discussed. We suggest that patients on the wait-list for a deceased human kidney graft who are unlikely to receive one due to long waiting times are those for whom kidney xenotransplantation might first be considered. The potential risk of infection, public attitudes to xenotransplantation, and ethical, regulatory, and financial aspects are briefly addressed.

Attitudes to Clinical Pig Kidney Xenotransplantation among Medical Providers and Patients.

In addition to governmental regulation and scientific advancements, the World Health Organization requires extensive review of local opinions before initiating clinical trials of xenotransplantation (XTx). The purpose of this study was to assess the attitudes of health care providers and patients regarding XTx. An anonymous Likert-scale survey regarding attitudes toward XTx was distributed to pre- and post-kidney transplant patients, nephrologists, transplant surgeons, and nurses ("providers"). Patient and provider responses were described and compared. Regression analysis using patients' responses was performed to identify factors associated with XTx acceptance. Eighty percent (32/40) of providers and 69% (113/163) of patients were agreeable to clinical XTx if the risks and results were likely to be similar to kidney allotransplantation (P<0.05). Kidney providers rated the influence of religious beliefs in medical decisions (45% versus 15%) and genetic engineering (43% versus 25%) as being more important than did patients (P<0.05). A small proportion in both groups (<15%) reported concerns about (1) potential personality changes, (2) how others would interact, (3) a perception of being "less human," or (4) morals or ethics. Logistic regression found that the odds of patients accepting XTx were greater if they had no religious concerns (OR, 25.10; 95% CI, 2.59 to 243.00), but acceptance was less likely if they were not willing to use XTx as a bridge to allotransplantation (OR, 0.18; 95% CI, 0.51 to 0.70). (1) If outcomes were similar to allotransplantation, XTx support was strong among both providers and patients; (2) providers overestimated the influence of religious beliefs and genetic engineering on patient medical decisions, although religious beliefs are associated with XTx acceptance; (3) XTx use as a bridge to allotransplant was associated with XTx acceptance; and (4) psychosocial concerns were low for either group. Future studies among other communities are warranted to assess if similar attitudes exist.

Clinical Pig Kidney Xenotransplantation: How Close Are We?

Patients with ESKD who would benefit from a kidney transplant face a critical and continuing shortage of kidneys from deceased human donors. As a result, such patients wait a median of 3.9 years to receive a donor kidney, by which time approximately 35% of transplant candidates have died while waiting or have been removed from the waiting list. Those of blood group B or O may experience a significantly longer waiting period. This problem could be resolved if kidneys from genetically engineered pigs offered an alternative with an acceptable clinical outcome. Attempts to accomplish this have followed two major paths: deletion of pig xenoantigens, as well as insertion of "protective" human transgenes to counter the human immune response. Pigs with up to nine genetic manipulations are now available. In nonhuman primates, administering novel agents that block the CD40/CD154 costimulation pathway, such as an anti-CD40 mAb, suppresses the adaptive immune response, leading to pig kidney graft survival of many months without features of rejection (experiments were terminated for infectious complications). In the absence of innate and adaptive immune responses, the transplanted pig kidneys have generally displayed excellent function. A clinical trial is anticipated within 2 years. We suggest that it would be ethical to offer a pig kidney transplant to selected patients who have a life expectancy shorter than the time it would take for them to obtain a kidney from a deceased human donor. In the future, the pigs will also be genetically engineered to control the adaptive immune response, thus enabling exogenous immunosuppressive therapy to be significantly reduced or eliminated.

Chronic dialysis in patients with end-stage renal disease: Relevance to kidney xenotransplantation.

Renal allotransplantation clearly offers better survival and quality of life for end-stage renal disease (ESRD) patients than chronic dialysis. The median waiting time for a deceased donor kidney in a suitable ESRD patient is 3.9years. The initial candidates for pig kidney xenotransplantation will be those with ESRD unlikely to receive an allograft within a reasonable period of time. It is thus reasonable to ascertain whether clinical trials of xenotransplantation might likewise offer superior outcomes. Chronic dialysis in patients with ESRD is associated with poor quality of life, significant morbidity, and relatively high mortality, with only 56% surviving 3years and 42% at 5years. However, a significant number of these patients, because of comorbidities, frailty, etc, would not be considered for renal allotransplantation and likely not for xenotransplantation. As genetically engineered pig kidneys have satisfactorily supported life in immunosuppressed nonhuman primates for many months or even more than a year, consideration in carefully selected patients could be given to pig kidney xenotransplantation. We suggest that, in order to give a patient the best possible outcome, the pig kidney could be transplanted pre-emptively (before dialysis is initiated). If it fails at any stage, the patient would then begin chronic dialysis and continue to await an allograft. The present (limited) evidence is that failure of a pig graft would not be detrimental to a subsequent allograft.

211 Pig kidney xenotransplantation in baboons: Persistent rejection despite an important immunosuppression protocol

211 Pig kidney xenotransplantation in baboons: Persistent rejection despite an important immunosuppression protocol