Controlling CAR-T cell activity and specificity with synthetic SparX adapters.

While conventional chimeric antigen-receptor (CAR)-T therapies have shown remarkable clinical activity in some settings, they can induce severe toxicities and are rarely curative. To address these challenges, we developed a controllable cell therapy where synthetic D-domain-containing proteins (soluble protein antigen-receptor X-linker [SparX]) bind one or more tumor antigens and mark those cells for elimination by genetically modified T cells (antigen-receptor complex [ARC]-T). The chimeric antigen receptor was engineered with a D-domain that specifically binds to the SparX protein via a unique TAG, derived from human alpha-fetoprotein. The interaction is mediated through an epitope on the TAG that is occluded in the native alpha-fetoprotein molecule. In vitro and in vivo data demonstrate that the activation and cytolytic activity of ARC-T cells is dependent on the dose of SparX protein and only occurs when ARC-T cells are engaged with SparX proteins bound to antigen-positive cells. ARC-T cell specificity was also redirected in vivo by changing SparX proteins that recognized different tumor antigens to combat inherent or acquired tumor heterogeneity. The ARC-SparX platform is designed to expand patient and physician access to cell therapy by controlling potential toxicities through SparX dosing regimens and enhancing tumor elimination through sequential or simultaneous administration of SparX proteins engineered to bind different tumor antigens.

Preclinical Efficacy of BCMA-Directed CAR T Cells Incorporating a Novel D Domain Antigen Recognition Domain.

Chimeric antigen receptor (CAR) T-cell therapies directed against B-cell maturation antigen (BCMA) have shown compelling clinical activity and manageable safety in subjects with relapsed and refractory multiple myeloma (RRMM). Prior reported CAR T cells have mostly used antibody fragments such as humanized or murine single-chain variable fragments or camelid heavy-chain antibody fragments as the antigen recognition motif. Herein, we describe the generation and preclinical evaluation of ddBCMA CAR, which uses a novel BCMA binding domain discovered from our D domain phage display libraries and incorporates a 4-1BB costimulatory motif and CD3-zeta T-cell activation domain. Preclinical in vitro studies of ddBCMA CAR T cells cocultured with BCMA-positive cell lines showed highly potent, dose-dependent measures of cytotoxicity, cytokine production, T-cell degranulation, and T-cell proliferation. In each assay, ddBCMA CAR performed as well as the BCMA-directed scFv-based C11D5.3 CAR. Furthermore, ddBCMA CAR T cells demonstrated in vivo tumor suppression in three disseminated BCMA-expressing tumor models in NSG-immunocompromised mice. On the basis of these promising preclinical data, CART-ddBCMA is being studied in a first-in-human phase I clinical study to assess the safety, pharmacokinetics, immunogenicity, efficacy, and duration of effect for patients with RRMM (NCT04155749).

Meta-analysis of interpersonal discrimination and health-related behaviors.

OBJECTIVE: This article comprehensively examines the relationship between interpersonal discrimination (IPD) and health-related behavior (HB), expanding upon Pascoe and Richman's (2009) meta-analysis and research synthesis. METHOD: One hundred and twenty one articles providing zero-order correlations (or information allowing their calculation) between perceptions of IPD and a variety of HB outcomes were coded and analyzed using a random-effects meta-analysis model. One hundred and fifty six articles examining this relationship using multivariate models were also coded and summarized within a research synthesis. Subanalyses were performed for articles examining smoking, alcohol use/abuse, substance use/abuse, sexual risk, and diet/eating behaviors. Potential mediators of the IPD-HB relationship were also tallied when available. RESULTS: Compared to the original analysis, results suggest an overall attenuated but stable relationship between IPD and HBs. CONCLUSIONS: The documented meta-analytic associations between perceptions of IPD and a variety of HB provide supportive evidence for one pathway through which IPD heightens risk for negative physical health outcomes among marginalized groups. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

Social Relationships, Wealth, and Cardiometabolic Risk: Evidence from a National Longitudinal Study of U.S. Older Adults.

Objectives: To investigate multiple dimensions of social relationships related to biomarkers of cardiometabolic health and how their associations vary by wealth in older adults. Methods: Growth curve models were used to investigate the longitudinal associations between measures of both positive and negative social relationships and cardiometabolic risk (CMR) over a 10-year period from 2006 to 2016 and the moderation of this association by wealth in the Health and Retirement Study (HRS). Results: Older adults with better social relationships had lower CMR on average. The protective effects of positive social relationships, however, waned at older ages, particularly for low-wealth individuals. Discussion: Our results suggest that good social relationships promote healthy aging by buffering against harmful cardiometabolic consequences of psychosocial stress, particularly among relatively wealthy individuals. Efforts to improve old age health would be more effective when focusing simultaneously on fostering social connections and boosting financial resources.

Adeno-Associated Virus-Induced Dorsal Root Ganglion Pathology.

The administration of adeno-associated virus (AAV) vectors to nonhuman primates (NHP) via the blood or cerebrospinal fluid (CSF) can lead to dorsal root ganglion (DRG) pathology. The pathology is minimal to moderate in most cases; clinically silent in affected animals; and characterized by mononuclear cell infiltrates, neuronal degeneration, and secondary axonopathy of central and peripheral axons on histopathological analysis. We aggregated data from 33 nonclinical studies in 256 NHP and performed a meta-analysis of the severity of DRG pathology to compare different routes of administration, dose, time course, study conduct, age of the animals, sex, capsid, promoter, capsid purification method, and transgene. DRG pathology was observed in 83% of NHP that were administered AAV through the CSF, and 32% of NHP that received an intravenous (IV) injection. We show that dose and age at injection significantly affected the severity whereas sex had no impact. DRG pathology was minimal at acute time points (i.e., <14 days), similar from one to 5 months post-injection, and was less severe after 6 months. Vector purification method had no impact, and all capsids and promoters that we tested resulted in some DRG pathology. The data presented here from five different capsids, five different promoters, and 20 different transgenes suggest that DRG pathology is almost universal after AAV gene therapy in nonclinical studies using NHP. None of the animals receiving a therapeutic transgene displayed any clinical signs. Incorporation of sensitive techniques such as nerve-conduction velocity testing can show alterations in a minority of animals that correlate with the severity of peripheral nerve axonopathy. Monitoring sensory neuropathies in human central nervous system and high-dose IV clinical studies seems prudent to determine the functional consequences of DRG pathology.

Translational Feasibility of Lumbar Puncture for Intrathecal AAV Administration.

Preclinical studies have demonstrated that a single injection of an adeno-associated virus (AAV) vector into the cerebrospinal fluid (CSF) can achieve widespread gene transfer throughout the central nervous system. Successfully translating this approach to humans requires identifying factors that influence AAV distribution in the CSF so that optimal parameters can be replicated in the clinic. In the context of developing a motor neuron-targeted gene therapy for spinal muscular atrophy, we conducted studies in nonhuman primates to evaluate the impact of injection volume on spinal cord transduction after AAV delivery via lumbar puncture. Lumbar injection of an AAVhu68 vector targeted motor neurons throughout the spinal cord, but only in juvenile nonhuman primates administered large injection volumes, equivalent to about half of the total CSF volume. Upon repeating this study with clinically relevant injection volumes and larger animals, we found that lumbar puncture failed to achieve significant transduction of the spinal cord. In contrast, vector administered into the cisterna magna distributed reproducibly throughout the spinal cord in both juvenile and adult animals. These findings highlight the challenges of translating AAV delivery via lumbar puncture to humans and suggest that delivery into the cisterna magna may represent a more feasible alternative.

Defining gentrification for epidemiologic research: A systematic review.

Neighborhoods have a profound impact on individual health. There is growing interest in the role of dynamic changes to neighborhoods-including gentrification-on the health of residents. However, research on the association between gentrification and health is limited, partly due to the numerous definitions used to define gentrification. This article presents a systematic review of the current state of literature describing the association between gentrification and health. In addition, it provides a novel framework for addressing important next steps in this research. A total of 1393 unique articles were identified, 122 abstracts were reviewed, and 36 articles published from 2007-2020 were included. Of the 36 articles, 9 were qualitative, 24 were quantitative, and 3 were review papers. There was no universally accepted definition of gentrification; definitions often used socioeconomic variables describing demographics, housing, education, and income. Health outcomes associated with gentrification included self-reported health, preterm birth, mental health conditions, alcohol use, psychosocial factors, and health care utilization, though the direction of this association varied. The results of this review also suggest that the impact of gentrification on health is not uniform across populations. For example, marginalized populations, such as Black residents and the elderly, were impacted more than White and younger residents. In addition, we identified multiples gaps in the research, including the need for a conceptual model, future mechanistic studies, and interventions.

Addressing health inequalities in diverse, rural communities: An unmet need.

Research on rural health needs to represent the diverse demographics of these regions by carefully considering the distinct characteristics, inequities, and stressors occurring in rural communities. Drawing from our own findings and other empirical investigations examining diverse rural communities, we propose several considerations to guide future endeavors toward more inclusive rural health research. These include population-health assessment tools that consider minority stress and intervention strategies designed to reflect both the environmental and socio-cultural contexts of rural residents.

Quantifying intersectionality: An important advancement for health inequality research.

BACKGROUND: Intersectionality is a powerful theoretical framework that is useful in describing the lived experiences of people with multiple marginalized statuses. By focusing on power and domination (e.g., racism, sexism), and the ways in which they are inextricably linked and mutually constructing, researchers can better understand experiences of all people, not just those with one or more master statuses. This framework is valuable in understanding how discrimination relates to health and in attempts to reduce health disparities. RATIONALE: Population health researchers have only recently begun to consider intersectionality in their theories and measurement (Bowleg, 2012), and have been hindered by the challenges of measuring and analyzing experiences of discrimination in intersectional ways. We need new methodological strategies to enable empirical research to catch up with theoretical advances. CONCLUSIONS: The pair of articles in this issue by Scheim and Bauer (2019), and Bauer and Scheim (2019), offer important new data collection instruments and data analytic strategies to advance our ability to measure discrimination intersectionally. When using these new tools, it is important to not lose track of the origins and historical underpinnings of intersectionality and to focus on the transformative goal of intersectionality to eradicate inequality.

AAV8 Gene Therapy for Crigler-Najjar Syndrome in Macaques Elicited Transgene T Cell Responses That Are Resident to the Liver.

Systemic delivery of adeno-associated viral (AAV) vectors has been evaluated for the treatment of several liver diseases, including homozygous familial hypercholesterolemia, ornithine transcarbamylase deficiency, and hemophilia. Here, we evaluated this approach for the treatment of Crigler-Najjar syndrome. We administered wild-type rhesus macaques with 1.0 × 1013 or 2.5 × 1013 genome copies/kg of an AAV serotype 8 vector expressing a codon-optimized version of human uridine diphosphate glucuronosyl transferase 1A1 (UGT1A1) from a liver-specific promoter. We extensively studied vector biodistribution, transgene expression, and immune responses following vector administration. All rhesus macaques survived until their scheduled necropsy at day 56 and showed no clinical abnormalities during the course of the study. Macaques administered with either vector dose developed a T cell response to the AAV capsid and/or transgene. We mapped the immunodominant epitope in the human UGT1A1 sequence, and we found no correlation between peripheral and tissue-resident lymphocyte responses. Upon further investigation, we characterized CD107a+, granzyme B+, CD4+, and CD8+ transgene-specific cellular responses that were restricted to tissue-resident T cells. This study highlights the importance of studying immune responses at the vector transduction site and the limited usefulness of blood as a surrogate to evaluate tissue-restricted T cell responses.

Determining the Minimally Effective Dose of a Clinical Candidate AAV Vector in a Mouse Model of Crigler-Najjar Syndrome.

Liver metabolism disorders are attractive targets for gene therapy, because low vector doses can reverse the buildup of toxic metabolites in the blood. Crigler-Najjar syndrome is an inherited disorder of bilirubin metabolism that is caused by the absence of uridine diphosphate glucuronosyl transferase 1A1 (UGT1A1) activity. This syndrome is characterized by hyperbilirubinemia and jaundice. Unfortunately, current phototherapy treatment is not effective long term. We intravenously injected phototherapy-rescued adult UGT1 knockout mice with 2.5 × 1010-2.5 × 1013 genome copies (GC)/kg of a clinical candidate vector, AAV8.TBG.hUGT1A1co, to study the treatment of disease compared to vehicle-only control mice. There were no apparent vector-related laboratory or clinical sequelae; the only abnormalities in clinical pathology were elevations in liver transaminases, primarily in male mice at the highest vector dose. Minimal to mild histopathological findings were present in control and vector-administered male mice. At vector doses greater than 2.5 × 1011 GC/kg, we observed a reversal of total bilirubin levels to wild-type levels. Based on a significant reduction in serum total bilirubin levels, we determined the minimally effective dose in this mouse model of Crigler-Najjar syndrome to be 2.5 × 1011 GC/kg.

Toxicology Study of Intra-Cisterna Magna Adeno-Associated Virus 9 Expressing Iduronate-2-Sulfatase in Rhesus Macaques.

Hunter syndrome is an X-linked recessive disease caused by deficiency of the lysosomal enzyme iduronate-2-sulfatase. The severe form of this progressive, systemic, and neurodegenerative disease results in loss of cognitive skills and early death. Several clinical trials are evaluating adeno-associated virus 9 for the treatment of neurodegenerative diseases using systemic or intrathecal lumbar administration. In large animals, administration via suboccipital puncture gives better brain transduction than lumbar administration. Here, we conducted a good laboratory practice-compliant investigational new drug-enabling study to determine the safety of suboccipital adeno-associated virus 9 gene transfer of human iduronate-2-sulfatase into nonhuman primates. Thirteen rhesus macaques received vehicle or one of two doses of vector with or without immunosuppression. We assessed in-life safety and immune responses. Animals were euthanized 90 days post-administration and sampled for histopathology and biodistribution. The procedure was well tolerated in all animals. Minimal mononuclear cerebrospinal fluid pleocytosis occurred in some animals. Asymptomatic minimal-to-moderate toxicity to some dorsal root ganglia sensory neurons and their associated axons occurred in all vector-treated animals. This study supports the clinical development of suboccipital adeno-associated virus 9 delivery for severe Hunter syndrome and highlights a potential toxicity that warrants monitoring in first-in-human studies.

Toxicology Study of Intra-Cisterna Magna Adeno-Associated Virus 9 Expressing Human Alpha-L-Iduronidase in Rhesus Macaques.

Mucopolysaccharidosis type I is a recessive genetic disease caused by deficiency of the lysosomal enzyme α-L-iduronidase, which leads to a neurodegenerative and systemic disease called Hurler syndrome in its most severe form. Several clinical trials are evaluating adeno-associated virus serotype 9 (AAV9) for the treatment of neurodegenerative diseases. Although these trials focus on systemic or lumbar administration, intrathecal administration via suboccipital puncture into the cisterna magna has demonstrated remarkable efficacy in large animals. We, therefore, conducted a good laboratory practice-compliant non-clinical study to investigate the safety of suboccipital AAV9 gene transfer of human α-L-iduronidase into nonhuman primates. We dosed 22 rhesus macaques, including three immunosuppressed animals, with vehicle or one of two doses of vector. We assessed in-life safety and immune responses. Animals were euthanized 14, 90, or 180 days post-vector administration and evaluated for histopathology and biodistribution. No procedure-related lesions or adverse events occurred. All vector-treated animals showed a dose-dependent mononuclear pleocytosis in the cerebrospinal fluid and minimal to moderate asymptomatic degeneration of dorsal root ganglia neurons and associated axons. These studies support the clinical development of suboccipital AAV delivery for Hurler syndrome and highlight a potential sensory neuron toxicity that warrants careful monitoring in first-in-human studies.

Intrathecal Viral Vector Delivery of Trastuzumab Prevents or Inhibits Tumor Growth of Human HER2-Positive Xenografts in Mice.

Breast cancer brain metastases are a deadly sequela of primary breast tumors that overexpress human epidermal growth factor receptor 2 (HER2); median survival for patients with these tumors is 10 to 13 months from the time of diagnosis. Current treatments for HER2-positive breast cancer brain metastases are invasive, toxic, and largely ineffective. Here, we have developed an adeno-associated virus serotype 9 (AAV9) vector to express the anti-HER2 monoclonal antibody trastuzumab (Herceptin) in vivo A single prophylactic intrathecal administration of AAV9.trastuzumab vector in a novel orthotopic Rag1-/- murine xenograft model of HER2-positive breast cancer brain metastases significantly increased median survival, attenuated brain tumor growth, and preserved both the HER2 antigen specificity and the natural killer cell-associated mechanism of action of trastuzumab. When administered as a tumor treatment, AAV9.trastuzumab increased median survival. Dose-escalation studies revealed that higher doses of AAV9.trastuzumab resulted in smaller tumor volumes. Our results indicate that intrathecal AAV9.trastuzumab may provide significant antitumor activity in patients with HER2-positive breast cancer brain metastases.Significance: Intrathecal delivery of trastuzumab via adeno-associated virus has the potential to become a novel, integral part of adjuvant therapy for patients with HER2-positive breast cancer brain metastases. Cancer Res; 78(21); 6171-82. ©2018 AACR.

Repairing the Leaky Pipeline: A Motivationally Supportive Intervention to Enhance Persistence in Undergraduate Science Pathways.

The current study reports on the efficacy of a multi-faceted motivationally designed undergraduate enrichment summer program for supporting science, technology, engineering and math (STEM) persistence. Structural equation modeling was used to compare summer program participants (n = 186), who participated in the program between their first and second years in college, to a propensity score matched comparison sample (n = 401). Participation in the summer program positively predicted science motivation (self-efficacy, task value), assessed eight months after the end of the program (second year in college). The summer enrichment program was also beneficial for science persistence variables, as evidenced by significant direct and indirect effects of the program on science course completion during students' third year of college and students' intentions to pursue a science research career assessed during the third year of college. In general, the program was equally beneficial for all participants, but ancillary analyses indicated added benefits with respect to task value for students with relatively lower prior science achievement during the first year of college and with respect to subsequent science course taking for males. Implications for developing effective interventions to reduce the flow of individuals out of STEM fields and for translating motivational theory into practice are discussed.

Severe Toxicity in Nonhuman Primates and Piglets Following High-Dose Intravenous Administration of an Adeno-Associated Virus Vector Expressing Human SMN.

Neurotropic adeno-associated virus (AAV) serotypes such as AAV9 have been demonstrated to transduce spinal alpha motor neurons when administered intravenously (i.v.) at high doses. This observation led to the recent successful application of i.v. AAV9 delivery to treat infants with spinal muscular atrophy, an inherited deficiency of the survival of motor neuron (SMN) protein characterized by selective death of lower motor neurons. To evaluate the efficiency of motor neuron transduction with an AAV9 variant (AAVhu68) using this approach, three juvenile nonhuman primates (NHPs; aged 14 months) and three piglets (aged 7-30 days) were treated with an i.v. injection of an AAVhu68 vector carrying a human SMN transgene at a dose similar to that employed in the spinal muscular atrophy clinical trial. Administration of 2 × 1014 genome copies per kilogram of body weight resulted in widespread transduction of spinal motor neurons in both species. However, severe toxicity occurred in both NHPs and piglets. All three NHPs exhibited marked transaminase elevations. In two NHPs, the transaminase elevations resolved without clinical sequelae, while one NHP developed acute liver failure and shock and was euthanized 4 days after vector injection. Degeneration of dorsal root ganglia sensory neurons was also observed, although NHPs exhibited no clinically apparent sensory deficits. There was no correlation between clinical findings and T-cell responses to the vector capsid or transgene product in NHPs. Piglets demonstrated no evidence of hepatic toxicity, but within 14 days of vector injection, all three animals exhibited proprioceptive deficits and ataxia, which profoundly impaired ambulation and necessitated euthanasia. These clinical findings correlated with more severe dorsal root ganglia sensory neuron lesions than those observed in NHPs. The liver and sensory neuron findings appear to be a direct consequence of AAV transduction independent of an immune response to the capsid or transgene product. The present results and those of another recent study utilizing a different AAV9 variant and transgene indicate that systemic and sensory neuron toxicity may be general properties of i.v. delivery of AAV vectors at high doses, irrespective of the capsid serotype or transgene. Preclinical and clinical studies involving high systemic doses of AAV vectors should include careful monitoring for similar toxicities.

Racial (vs. self) affirmation as a protective mechanism against the effects of racial exclusion on negative affect and substance use vulnerability among black young adults.

Affirming one's racial identity may help protect against the harmful effects of racial exclusion on substance use cognitions. This study examined whether racial versus self-affirmation (vs. no affirmation) buffers against the effects of racial exclusion on substance use willingness and substance use word associations in Black young adults. It also examined anger as a potential mediator of these effects. After being included, or racially excluded by White peers, participants were assigned to a writing task: self-affirmation, racial-affirmation, or describing their sleep routine (neutral). Racial exclusion predicted greater perceived discrimination and anger. Excluded participants who engaged in racial-affirmation reported reduced perceived discrimination, anger, and fewer substance use cognitions compared to the neutral writing group. This relation between racial-affirmation and lower substance use willingness was mediated by reduced perceived discrimination and anger. Findings suggest racial-affirmation is protective against racial exclusion and, more generally, that ethnic based approaches to minority substance use prevention may have particular potential.

Validation of the Food-Linked Virtual Response task.

This research validates a computerized dietary selection task (Food-Linked Virtual Response or FLVR) for use in studies of food consumption. In two studies, FLVR task responses were compared with measures of health consciousness, mood, body mass index, personality, cognitive restraint toward food, and actual food selections from a buffet table. The FLVR task was associated with variables which typically predict healthy decision-making and was unrelated to mood or body mass index. Furthermore, the FLVR task predicted participants' unhealthy selections from the buffet, but not overall amount of food. The FLVR task is an inexpensive, valid, and easily administered option for assessing momentary dietary decisions.

Heteronormativity and practitioner-patient interaction.

Heteronormativity is the presumption of heterosexuality as the default sexual orientation and can result in discrimination against the lesbian, gay, and bisexual (LGB) population. This study serves as one of the first experimental studies to examine heteronormative perceptions in communication and their effects on practitioner-patient relationships. LGB participants were randomly assigned to read either heteronormative or non-heteronormative vignettes of a doctor-patient interaction. They then indicated how much health-relevant information they would disclose to the doctor in the vignette and their level of trust in the doctor. In the heteronormative condition, participants were less likely to disclose health-relevant information to the doctor in the vignette and were less trustful of the doctor as compared to those in the non-heteronormative condition. These results have important health implications, as lack of disclosure and trust may prevent people from getting needed care and prevent doctors from giving the best health advice possible. The results of this study provide further evidence that there is a need for more education for all health care professionals to feel comfortable while respectfully communicating with and treating patients who do not identify as heterosexual in order to ensure the best health care experience.