HIF2 Inactivation and Tumor Suppression with a Tumor-Directed RNA-Silencing Drug in Mice and Humans.

PURPOSE: HIF2α is a key driver of kidney cancer. Using a belzutifan analogue (PT2399), we previously showed in tumorgrafts (TG) that ∼50% of clear cell renal cell carcinomas (ccRCC) are HIF2α dependent. However, prolonged treatment induced resistance mutations, which we also identified in humans. Here, we evaluated a tumor-directed, systemically delivered, siRNA drug (siHIF2) active against wild-type and resistant-mutant HIF2α. EXPERIMENTAL DESIGN: Using our credentialed TG platform, we performed pharmacokinetic and pharmacodynamic analyses evaluating uptake, HIF2α silencing, target gene inactivation, and antitumor activity. Orthogonal RNA-sequencing studies of siHIF2 and PT2399 were pursued to define the HIF2 transcriptome. Analyses were extended to a TG line generated from a study biopsy of a siHIF2 phase I clinical trial (NCT04169711) participant and the corresponding patient, an extensively pretreated individual with rapidly progressive ccRCC and paraneoplastic polycythemia likely evidencing a HIF2 dependency. RESULTS: siHIF2 was taken up by ccRCC TGs, effectively depleted HIF2α, deactivated orthogonally defined effector pathways (including Myc and novel E2F pathways), downregulated cell cycle genes, and inhibited tumor growth. Effects on the study subject TG mimicked those in the patient, where HIF2α was silenced in tumor biopsies, circulating erythropoietin was downregulated, polycythemia was suppressed, and a partial response was induced. CONCLUSIONS: To our knowledge, this is the first example of functional inactivation of an oncoprotein and tumor suppression with a systemic, tumor-directed, RNA-silencing drug. These studies provide a proof-of-principle of HIF2α inhibition by RNA-targeting drugs in ccRCC and establish a paradigm for tumor-directed RNA-based therapeutics in cancer.

The hospital autopsy: the importance in keeping autopsy an option.

Autopsy has been one of the most powerful diagnostic tools in medicine for over a century. Despite its importance in establishing cause of death and elucidating pathophysiology of disease, rates of hospital autopsies continue to decline. In this study we aim to determine if physicians believe autopsies are essential to patient care through discussion of autopsy with families. At the same time, we analyzed whether families are more willing to consent to autopsy if physicians are involved in autopsy discussion at the time of death, and what may be the reasons for not wanting an autopsy. Our results showed a doubling in autopsy consent when autopsy was discussed by the physician. Additionally, the biggest reason for families not consenting to autopsy was because they believed they already knew what caused death. The emergence of Coronavirus 2019 (COVID-19) has re-established the value of autopsy, as seen by increased autopsy rates in the past year. This study demonstrates that physician conversation with families on autopsy leads to an increased chance of autopsy consent.

Impact of mobilization regimen on multiday collection of peripheral blood CD34+ cells by large volume leukapheresis.

BACKGROUND: The collection yield of hematopoietic progenitors cell (HPC) by leukapheresis is critical for a successful transplantation, which often requires multiday collections to achieve the collection goal. STUDY DESIGN AND METHODS: Collection procedures of 181 patients who underwent leukapheresis for more than 1 day were reviewed. Patients were separated into six groups based on the mobilization regimen: G-CSF on day 1 (D1) and day 2 (D2) (G-G); G-CSF on D1 and G-CSF and plerixafor on D2 (G-GP); G-CSF and plerixafor on day D1 and D2 (GP-GP); G-CSF and plerixafor on D1 and G-CSF on D2 (GP-G); chemotherapy and G-CSF on D1 and D2 (GC-GC); and chemotherapy, G-CSF, and plerixafor on D1 and D2 (GCP-GCP). Patient's pre-collection CD34 count (pre-CD34) on D1 and D2 were compared in the same individual and among groups. RESULTS: We found D2 pre-CD34 were significantly decreased in G-G, GP-G, and GP-GP groups and significantly increased in G-GP group (p < .001) using a repeated measures ANOVA analysis. D2 pre-CD34 remained at similar levels as D1 in GC-GC and GCP-GCP groups. A multiple regression analysis showed that the mobilization regimen was the only factor that significantly affected pre-CD34 D2/D1 ratio (p < .001). There was a significant difference in the pre-CD34 D2/D1 ratio (p < .001) among these six groups with the lowest in GP-G group (0.40 ± 0.45), and the highest in G-GP group (2.35 ± 0.36). DISCUSSION: Mobilization regimen has significant impact on pre-collection CD34 count. Apheresis facilities may change mobilizing drugs accordingly to achieve a specific HPC goal.

Rescue Hepatectomy and Anhepatic Phase Management After Primary Nonfunction in a Liver Transplant.

Primary nonfunction is a rare but lethal complication that occurs in a small number of liver transplants. When primary nonfunction occurs, the only definite treatment is retransplant; however, another liver might not be readily available at that time. Hence, a surgeon should be aware of the various options available at hand for patient care during the time interval between the primary nonfunction and retransplant. Here, we describe the management strategy that was devised to take care of an unstable anhepatic patient in the intensive care unit, care of the patient during anhepatic phase, and successful outcome with a second liver transplant. Our index patient was a recipient of a liver donated after cardiac death. While in the operating room, after reperfusion of the liver, the patient had right heart dysfunction leading to hemodynamic instability and congestion of the liver, which culminated in primary nonfunction. Graft hepatectomy had to be done on postoperative day 1 because of deteriorating condition of the patient, and the patient was maintained in anhepatic phase in the intensive care unit for 27 hours.

The hospital autopsy: the importance in keeping autopsy an option

Autopsy has been one of the most powerful diagnostic tools in medicine for over a century. Despite its importance in establishing cause of death and elucidating pathophysiology of disease, rates of hospital autopsies continue to decline. In this study we aim to determine if physicians believe autopsies are essential to patient care through discussion of autopsy with families. At the same time, we analyzed whether families are more willing to consent to autopsy if physicians are involved in autopsy discussion at the time of death, and what may be the reasons for not wanting an autopsy. Our results showed a doubling in autopsy consent when autopsy was discussed by the physician. Additionally, the biggest reason for families not consenting to autopsy was because they believed they already knew what caused death. The emergence of Coronavirus 2019 (COVID-19) has re-established the value of autopsy, as seen by increased autopsy rates in the past year. This study demonstrates that physician conversation with families on autopsy leads to an increased chance of autopsy consent.

Digital Pathology Initiatives and Experience of a Large Academic Institution During the Coronavirus Disease 2019 (COVID-19) Pandemic.

CONTEXT.­: Pathology practices have begun integrating digital pathology tools into their routine workflow. During 2020, the coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), emerged as a pandemic, causing a global health crisis that significantly affected the world population in several areas, including medical practice, and pathology was no exception. OBJECTIVE.­: To summarize our experience in implementing digital pathology for remote primary diagnosis, education, and research during this pandemic. DESIGN.­: We surveyed our pathologists (all subspecialized) and trainees to gather information about their use of digital pathology tools before and during the pandemic. Quality assurance and slide distribution data were also examined. RESULTS.­: During the pandemic, the widespread use of digital tools in our institution allowed a smooth transition of most clinical and academic activities into remote with no major disruptions. The number of pathologists using whole slide imaging (WSI) for primary diagnosis increased from 20 (62.5%) to 29 (90.6%) of a total of 32 pathologists, excluding renal pathology and hematopathology, during the pandemic. Furthermore, the number of pathologists exclusively using whole slide imaging for primary diagnosis also increased from 2 (6.3%) to 5 (15.6%) during the pandemic. In 35 (100%) survey responses from attending pathologists, 21 (60%) reported using whole slide imaging for remote primary diagnosis following the Centers for Medicare and Medicaid Services waiver. Of these 21 pathologists, 18 (86%) responded that if allowed, they will continue using whole slide imaging for remote primary diagnosis after the pandemic. CONCLUSIONS.­: The pandemic served as a catalyst to pathologists adopting a digital workflow into their daily practice and realizing the logistic and technical advantages of such tools.

Acute fatal myocarditis after a single dose of anti-PD-1 immunotherapy, autopsy findings: a case report.

Nivolumab (PD-1 inhibitor) and Ipilimumab (CTLA-34 inhibitor) are both commonly used immune checkpoint inhibitor therapies for various cancers. Various adverse events are associated with these therapies, including hepatitis, nephritis, dermatitis, and myocarditis. It is believed these adverse events occur in part because modified cellular receptors lead to enhanced CD4 and CD8 lymphoproliferation. These events usually occur after several months and rounds of treatment. Here we present a case of an 81-year-old male with recurrent renal cell carcinoma (RCC) who experienced myocarditis after only a single dose of combination therapy with Nivolumab and Ipilimumab. He presented with elevated troponins and a third-degree heart block; three days after admission he died. Histologic examination revealed a predominance of CD3 T cells (CD4 > CD8) and CD68 macrophages, with occasional CD20 B cells. C4d staining was negative in the interstitial capillaries, suggesting that antibody-mediated injury of endothelial cells did not play a significant role in the pathogenesis of this myocarditis. Additional studies ruled out an infectious etiology. Immune checkpoint inhibitors are increasingly more common, and it is important clinicians are aware patients can present with myocarditis early in the course of treatment.