Wednesday, December 31, 2025

RORA neurodevelopmental disorder

Inspired by a patient

Talarico M, de Bellescize J, De Wachter M, Le Guillou X, Le Meur G, Egloff M, Isidor B, Cogné B, Beysen D, Rollier P, Fradin M, Pasquier L, Guella I, Hickey SE, Benke PJ, Shillington A, Kumps C, Vanakker O, Gerkes EH, Lakhani S, Romanova I, Kanivets I, Brugger M, Vill K, Caylor RC, Skinner C, Tinker RJ, Stödberg T, Nümann A, Haack TB, Deininger N, Hengel H, Jury J, Conrad S, Mercier S, Yoon G, Tsuboyama M, Barcia G, Gitiaux C, Rio M, Bevot A, Redon S, Uguen K, Wonneberger A, Schulz A, Timmann D, Karlowicz DH, Chatron N, Carnevale A, Mahida S, Õunap K, Kury S, Cabet S, Lesca G. RORA-neurodevelopmental disorder: A unique triad of developmental disabilities, cerebellar anomalies, and myoclonic seizures. Genet Med. 2025 Apr;27(4):101347. doi: 10.1016/j.gim.2024.101347. Epub 2024 Dec 17. PMID: 39707840.

Abstract

Purpose: RORA encodes the RAR-related orphan receptor-α, playing a pivotal role in cerebellar maturation and function. Here, we report the largest series of individuals with RORA-related-neurodevelopmental disorder.

Methods: Forty individuals (30 unrelated; 10 siblings from 4 families) carrying RORA pathogenic/likely pathogenic variants were collected through an international collaboration.

Results: The 33 variants (29 de novo, 4 inherited, and 1 shared), identified by genome/exome sequencing (n = 21), chromosomal microarray analysis (n = 7), or gene panels (n = 4), included frameshift (n = 18/33), missense (n = 9/33), and stop codon (n = 6/33). Developmental disability (n = 32/37), intellectual disability (n = 22/32), and cerebellar signs (n = 25/34) were the most striking clinical features. Cerebellar symptoms were divided into early-onset, late-onset, and progressive subgroups. Cerebellar hypoplasia, atrophy, or both (n = 16/25) were more frequent in individuals with missense variants in the DNA-binding domain. Epilepsy (n = 18/38), with prominent myoclonic seizure types (n = 11/18), was classified in (1) genetic generalized epilepsy (n = 10/18) with a syndromic diagnosis identifiable for 6: epilepsy with eyelid myoclonia (n = 5/6) and epilepsy with myoclonic absence (n = 1/6); (2) developmental and epileptic encephalopathy (n = 5/18); and (3) unclassified (n = 3/18). A participant with rapid deterioration of visual acuity and cone/rod dystrophy was reported.

Conclusion: Missense variants in DNA-binding domain correlate to a more severe cerebellar phenotype. The RORA-related-neurodevelopmental disorder triad comprises developmental disability, cerebellar features, and a spectrum of myoclonic epilepsy.

Guissart C, Latypova X, Rollier P, Khan TN, Stamberger H, McWalter K, Cho MT, Kjaergaard S, Weckhuysen S, Lesca G, Besnard T, Õunap K, Schema L, Chiocchetti AG, McDonald M, de Bellescize J, Vincent M, Van Esch H, Sattler S, Forghani I, Thiffault I, Freitag CM, Barbouth DS, Cadieux-Dion M, Willaert R, Guillen Sacoto MJ, Safina NP, Dubourg C, Grote L, Carré W, Saunders C, Pajusalu S, Farrow E, Boland A, Karlowicz DH, Deleuze JF, Wojcik MH, Pressman R, Isidor B, Vogels A, Van Paesschen W, Al-Gazali L, Al Shamsi AM, Claustres M, Pujol A, Sanders SJ, Rivier F, Leboucq N, Cogné B, Sasorith S, Sanlaville D, Retterer K, Odent S, Katsanis N, Bézieau S, Koenig M, Davis EE, Pasquier L, Küry S. Dual Molecular Effects of Dominant RORA Mutations Cause Two Variants of Syndromic Intellectual Disability with Either Autism or Cerebellar Ataxia. Am J Hum Genet. 2018 May 3;102(5):744-759. doi: 10.1016/j.ajhg.2018.02.021. Epub 2018 Apr 12. PMID: 29656859; PMCID: PMC5986661.

Abstract

RORα, the RAR-related orphan nuclear receptor alpha, is essential for cerebellar development. The spontaneous mutant mouse staggerer, with an ataxic gait caused by neurodegeneration of cerebellar Purkinje cells, was discovered two decades ago to result from homozygous intragenic Rora deletions. However, RORA mutations were hitherto undocumented in humans. Through a multi-centric collaboration, we identified three copy-number variant deletions (two de novo and one dominantly inherited in three generations), one de novo disrupting duplication, and nine de novo point mutations (three truncating, one canonical splice site, and five missense mutations) involving RORA in 16 individuals from 13 families with variable neurodevelopmental delay and intellectual disability (ID)-associated autistic features, cerebellar ataxia, and epilepsy. Consistent with the human and mouse data, disruption of the D. rerio ortholog, roraa, causes significant reduction in the size of the developing cerebellum. Systematic in vivo complementation studies showed that, whereas wild-type human RORA mRNA could complement the cerebellar pathology, missense variants had two distinct pathogenic mechanisms of either haploinsufficiency or a dominant toxic effect according to their localization in the ligand-binding or DNA-binding domains, respectively. This dichotomous direction of effect is likely relevant to the phenotype in humans: individuals with loss-of-function variants leading to haploinsufficiency show ID with autistic features, while individuals with de novo dominant toxic variants present with ID, ataxia, and cerebellar atrophy. Our combined genetic and functional data highlight the complex mutational landscape at the human RORA locus and suggest that dual mutational effects likely determine phenotypic outcome.

Sayad A, Noroozi R, Omrani MD, Taheri M, Ghafouri-Fard S. Retinoic acid-related orphan receptor alpha (RORA) variants are associated with autism spectrum disorder. Metab Brain Dis. 2017 Oct;32(5):1595-1601. doi: 10.1007/s11011-017-0049-6. Epub 2017 Jun 12. PMID: 28608249.

Abstract

Autism spectrum disorder (ASD) is a neurodevelopmental disorder with various epidemiologic, genetic, epigenetic, and environmental factors being associated with it. The observed sex bias in ASD towards male has prompted investigators to propose sex-dependent mechanisms for ASD. Retinoic acid-related orphan receptor-alpha (RORA) is a new autism candidate gene that has been shown to be differentially regulated by male and female hormones. Previous studies have shown deregulation of its expression in the prefrontal cortex and the cerebellum of ASD patients. In the present study we aimed at identification of the possible associations between two functional polymorphisms in the RORA gene (rs11639084 and rs4774388) and the risk of ASD in 518 Iranian ASD patients and 472 age, gender, and ethnic-matched healthy controls by means of tetra primer-amplification refractory mutation system-PCR. The allele and genotype frequencies of rs11639084 were not significantly different between patients and controls. However, the allele frequencies of rs4774388 showed significant overrepresentation of T allele in patients compared with controls (P = 0.04, OR (95% CI) =1.21 (1.01-1.46)). The rs4774388-TT genotype was significantly higher in patients compared with controls and was associated with ASD risk in dominant inheritance model (P = 0.04, OR (95% CI) =0.77 (0.59-0.99)). Haplotype analysis showed significant association of two estimated blocks of rs11639084/ rs4774388 with ASD risk. Consequently, the present data provide further evidence for RORA participation in the pathogenesis of ASD.

Monday, December 29, 2025

Treatment approaches in pediatric MOG antibody-associated disease

Suntornlohanakul R, Yea C, Yeh EA. Exploring Treatment Approaches in Pediatric MOG Antibody-Associated Disease: A Survey of Neurologists. Neurol Clin Pract. 2025 Aug;15(4):e200504. doi: 10.1212/CPJ.0000000000200504. Epub 2025 Jun 27. PMID: 40584638; PMCID: PMC12204770.

Abstract

Background and objectives: Diagnostic criteria for anti-myelin oligodendrocyte glycoprotein (MOG) antibody-associated diseases (MOGADs) were published in 2023, but debate regarding optimal therapeutic strategies for pediatric MOGAD exists. The aim of this study was to evaluate treatment approaches and preferred diagnostic investigations for pediatric MOGAD among neurologists.

Methods: A survey questionnaire focused on pediatric MOGAD treatment was launched through the Practice Current Section of Neurology® Clinical Practice in April-October 2024. Responses from neurologists were solicited through advertisements on American Academy of Neurology (AAN) social media platforms, on the AAN website, and in print editions of Neurology®, as well as through QR codes shared at professional neurologic meetings. The questionnaire included 12 questions evaluating clinical decision making after a first and second neuroinflammatory episode, in a child testing positive for MOG-IgG antibody. Demographic questions were included. Responses were evaluated using descriptive statistics. A comparative analysis was conducted between those who self-identified as neuroimmunologists (NIs) and those who did not.

Results: A total of 346 neurologists completed the survey (52.3% of general neurologists, 32.1% of NIs, and 15.6% in other neurology fields). Of all respondents, 90.5% chose to send serum MOG-IgG antibody after the first event (59.7% serum, 36.4% CSF + serum). For acute treatment, 84.1% chose to give a 3-5-day course of high-dose IV steroids. Approaches to steroid tapering varied, with 33.0% choosing a 2-4-week taper, 27.2% choosing a 7-12-week taper, and 21% not offering a steroid taper. 56.6% of non-NIs chose to initiate maintenance therapy after the first episode while only 18.9% of NIs chose to do so. After the second episode, 98.3% of all respondents recommended starting maintenance therapy, with rituximab (RTX) (37.1%) being the most frequently chosen agent, followed by monthly IV immunoglobulin (IVIG) (25.6%) and azathioprine (17.1%). NIs selected monthly IVIG (50%) over RTX (27.3%). The duration of treatment in relapsing cases varied: 42.9% elected to maintain treatment for 2 years or less and 35.3% for more than 2 years, and 21.8% chose to continue treatment indefinitely.

Discussion: The survey demonstrated substantial variability in management decisions related to MOGAD among neurologists, reflecting current gaps in knowledge about therapies for MOGAD. Future efforts are needed to improve the uptake of knowledge and ensure that current guidelines are effectively translated into clinical practice.

Rasmussen syndrome atypicalities

Caraballo RH, Valenzuela GR, Pociecha J, Princich JP, Gutierrez R, Beltran L, Lubieniecki F, Bartuluchi M. Rasmussen syndrome: an atypical presentation in ten patients. Epileptic Disord. 2018 Dec 1;20(6):468-478. doi: 10.1684/epd.2018.1007. PMID: 30530407.

Abstract

The aim of this study was to analyse the electroclinical and imaging findings and outcome of patients with Rasmussen syndrome (RS) with atypical manifestations. We conducted a retrospective, descriptive study of 10 of 44 consecutive patients with RS with atypical features, followed between 1999 and 2017. Six patients were boys and four were girls. The mean and median ages at onset of the seizures were 8.8 and 6.5 years, respectively (range: 4.6-13 years). All of the patients except one had seizures. Eight patients (80%) had epilepsia partialis continua that started at a mean age of 7.5 years (range: 7-15 years). In our series, hemiparesis without seizures was the first manifestation in three patients, one of whom had dual pathology. In two patients, the first manifestation was dyskinetic movements, followed by delayed-onset seizures associated with unilateral caudate atrophy. Two patients had a focal lesion mimicking focal cortical dysplasia as the first MRI abnormality; one of these two patients had epileptic spasms in clusters. Bilateral cerebral hemisphere involvement was observed in three patients during the course of the disease. Six of eight patients responded well to surgical treatment. Progressive hemiparesis alone or with delayed-onset seizures, dyskinetic movements associated with seizures, a focal lesion mimicking focal cortical dysplasia, and bilateral brain involvement were the atypical features recognized. Our series of patients responded well to surgery. Clinical, video-EEG, and neuroradiological follow-up is important for early confirmation of RS in order to initiate adequate management of the condition.

Olson HE, Lechpammer M, Prabhu SP, Ciarlini PD, Poduri A, Gooty VD, Anjum MW, Gorman MP, Loddenkemper T. Clinical application and evaluation of the Bien diagnostic criteria for Rasmussen encephalitis. Epilepsia. 2013 Oct;54(10):1753-60. doi: 10.1111/epi.12334. Epub 2013 Aug 23. PMID: 23980696.

Abstract

Purpose: The 2005 diagnostic criteria for Rasmussen encephalitis (RE) are based on seizures, clinical deficits, electroencephalography (EEG), neuroimaging, and pathology (Brain, 128, 2005, 451). We applied these criteria to patients evaluated for RE and epilepsy surgery controls to determine the sensitivity, specificity, and positive and negative predictive values (PPVs, NPVs) using pathology as the gold standard.

Methods: We identified patients evaluated for RE based on medical records from 1993 to 2011. Fifty-two control patients with refractory epilepsy, unilateral magnetic resonance imaging (MRI) changes, and biopsies were selected from an epilepsy surgery database from matching years. Patients meeting all three of group A and/or two of three group B criteria were classified as meeting full criteria (positive). Patients not meeting full criteria were classified as negative. When available, pathology findings were re-reviewed with neuropathologists, and MRI imaging was re-reviewed with a neuroradiologist.

Key findings: RE was considered in the differential diagnosis for 82 patients, of whom 35 had biopsies. Twenty patients met full criteria (positive) without another explanation, including seven for whom biopsy was required to meet criteria and one in whom another etiology was identified. Two patients met full criteria but had another explanation. Thirty-five met partial criteria (negative), of whom 14 had another etiology identified. Twenty-five met no criteria (negative). The diagnostic criteria had a sensitivity of 81% with four false negatives (criteria-negative, biopsy-positive) when compared to pathology as a gold standard. Five false positives (criteria positive, biopsy negative) had identifiable alternate diagnoses.

Significance: The 2005 Bien clinical diagnostic criteria for RE have reasonably high sensitivity and specificity and good clinical-pathologic correlation in most cases. We suggest modification of the criteria to allow inclusion of cases with well-described but less common features. Specifically we suggest making the diagnosis in the absence of epilepsia partialis continua (EPC) or clear progression of focal cortical deficits or MRI findings if biopsy is positive and two of the A criteria are met (B3 plus two of three A criteria). This would improve the sensitivity of the criteria.

Tuesday, December 16, 2025

Transplantation priorities

Courtesy of a colleague

Heart transplant patients in the United States typically spend months waiting for a donated organ. But Kayoko Hira was not a typical patient.

Mrs. Hira, the wife of a hotel magnate in Japan, flew to the United States in September 2021, went to the University of Chicago Medical Center and, within days, got a new heart from an American teenager who had died.

Soon after, The New York Times found, a charity run by her husband made a donation to a nonprofit group led by the heart surgeon’s wife. It was the only time the charity has ever given money to an American institution, according to its website.

More than 100,000 people in the United States are in need of a transplant, and each year thousands die waiting. But despite the shortage of organs, some American hospitals are aggressively courting international transplant patients, a New York Times investigation found.

They have advertised abroad, promoting short wait times and concierge services, particularly to patients in the Middle East, where about two-thirds of overseas transplant recipients are from. Several hospitals have signed contracts with foreign governments, setting prices for different organ transplants.

An international transplant patient can bring in as much as $2 million — far more than a U.S. patient paying through private insurance or a public program like Medicare.

In the past dozen years, more than 1,400 patients from abroad received a transplant in the United States after traveling specifically for the procedure. That was a small fraction of all U.S. transplants, and most transplant centers did not operate on international patients at all.

But The Times found that a handful of hospitals are increasingly catering to overseas patients, who make up an ever-larger share of their organ recipients: 11 percent for hearts and lungs at the University of Chicago; 20 percent for lungs at Montefiore Medical Center in the Bronx; 16 percent for lungs at UC San Diego Health; 10 percent for intestines at MedStar Georgetown University Hospital in Washington; and 8 percent for livers at Memorial Hermann-Texas Medical Center in Houston.

In many countries, this would be illegal. World leaders agreed in 2008 to fight so-called transplant tourism, and most nations do not provide organs to overseas patients. Yet the United States has long allowed it. The policy has drawn criticism in the past, such as when organs went to Saudi royals and a Japanese crime boss.

The Times analyzed every transplant performed in the United States since 2013. Overall, it found that patients who traveled from other countries received transplants faster than patients from America and were less likely to die waiting for an organ.

Dr. Mark Fox, a former chair of the transplant system’s ethics committee, said the findings were troubling, especially because overseas patients do not contribute to America’s pool of donated organs. “The unfortunate reality is that we don’t have enough organs,” he said. “When people jet in, get an organ and jet home, it’s a problem. It’s not fair.”

In response to requests for comment, several hospitals emphasized that giving transplants to international patients is legal, and said those patients do not get preferential treatment. Some described their work as humanitarian.

The transplant system used to effectively cap the number of transplants for overseas patients, focusing mostly on rare cases of particular need. Then, The Times found, things changed.

Irene Gebrael was an Arabic interpreter at Montefiore when, she said, she had an idea: If the hospital recruited international patients, it could serve a broader population — and make a lot of money.

“The hospital is always happy about the international rates,” Ms. Gebrael said in an interview. “Trust me.”

In 2013, she said, she persuaded hospital executives to start an international office. It was good timing for the transplant program.

For decades, hospitals that gave more than 5 percent of any organ type to “noncitizen nonresidents” could be audited and possibly penalized. But leaders of the transplant system ended that in 2012, concerned that it could lead to discrimination against undocumented immigrants, a group that commonly donated organs. (The Times analysis did not include patients who traveled to the United States for reasons other than transplants, such as immigrants.)

Since then, Montefiore, a modest transplant program in a low-income section of New York City, has increasingly treated international patients, which Ms. Gebrael said generated tens of millions of dollars in revenue. From 2020 to 2024, it provided organs to 49 such patients, the third most of any U.S. hospital.

“You make the hospital gain money, and the patients are happy, the embassies are satisfied,” said Ms. Gebrael, who left Montefiore in March and started her own business coordinating health care for overseas patients.

In a statement, Montefiore said it “strictly adheres” to all transplant system policies.

Since the restriction on international patients was lifted, only about 25 of America’s roughly 250 transplant centers have given more than one organ per year, on average, to overseas patients, The Times found.

At the hospitals that have provided the most transplants in recent years, nearly all of the international recipients came from Kuwait, Saudi Arabia, Israel, the United Arab Emirates and Qatar.

U.S. hospitals have advertised in Arabic-language magazines, hosted conferences in the Middle East and hired consultants there. “We are a world leader in transplant,” said a University of Chicago video posted with Arabic subtitles. It promised services including airport pickups and lodging assistance.

The University of Chicago performed transplants for 61 international patients from 2020 through 2024 — more than any other center.

The hospital declined to answer questions about treating overseas patients. It issued a statement saying it had “significantly improved access to organ transplantation in recent years, driven by our commitment to the health and well-being of the diverse and underserved populations of Chicago’s South Side and the broader region.”

Hospitals have competed to win over Middle Eastern governments, which often pay to send citizens abroad for care. Under federal rules, hospitals are supposed to consider each transplant patient individually and cannot strike broader agreements with governments. But at least three hospitals — the University of Chicago, Montefiore and Memorial Hermann — have signed such deals, according to former employees.

Those hospitals did not respond to questions about the contracts. A spokeswoman for UC San Diego said it had arrangements with Israeli health insurers. Data shows that it gave organs to 37 Israelis from 2020 to 2024, more than all other centers combined.

UC San Diego said international patients are referred there when they are especially high-risk or cannot get procedures in their home countries.

Dr. Mustafa Al-Mousawi, president of the Kuwait Transplant Society, said his government had agreements with multiple American hospitals. But because the government had to pay more than $1 million for each transplant, it did not send over everyone in need. The wealthy, he said, tend to have better access to the officials deciding who gets to go.

“This is unfortunate,” he said, “because there are many poor people in Kuwait who need transplants, too.”

The transplant system is designed to prioritize fairness: All potential recipients must join a national registry, and organs from deceased donors are distributed to patients based on factors such as how sick they are and how long they have been waiting.

But despite the safeguards, international patients have gained advantages.

In interviews, 25 current and former employees of eight hospitals said they had seen overseas patients receive preferential treatment.

To join the registry, patients must be certified by a hospital that they can survive a transplant and care for a new organ. But non-American patients are often fast-tracked, said workers who have evaluated potential organ recipients at the University of Chicago, Montefiore, UC San Diego and Georgetown. Some said they had been overruled by superiors after rejecting an international patient who smoked, had a high body mass index or otherwise did not meet criteria.

In text messages obtained by The Times, workers at one of those hospitals discussed how an overseas patient would inevitably make it onto the registry: “We will do him,” one said, “he is international 🤑.”

The hospitals said they fairly considered all patients to ensure they qualified for a transplant. “Our medical center doesn’t expedite listing evaluations for any patient,” the University of Chicago said.

When organs are retrieved from deceased donors in the United States, they are offered to potential recipients in priority order, as determined by algorithms.

But hospitals can request an “exception” to raise a patient’s priority level if they believe the algorithm does not fully capture the state of the person’s health. These requests go to committees of doctors where they are usually approved without much scrutiny, according to interviews with three physicians who do reviews.

In recent years, international patients who received lung or liver transplants were more likely than Americans to have gotten exceptions. For lung transplants, the data shows, 23 percent of overseas recipients got an exception; less than 5 percent of the Americans did. The rates were about even for hearts, the only other organ that allows for exceptions.

Kyella Fonseca, who worked as a financial coordinator for transplants at Montefiore from 2020 to 2022, said she had seen preference given to international patients, who she said generally paid more. She said colleagues sometimes altered or omitted health information in their medical records to help them get organs more quickly.

“We had patients who we’d been working with, who had been waiting their turn, and then someone from Kuwait would come and jump the line,” she said.

In its statement, Montefiore said, “The suggestion that we would compromise the health and well-being of any U.S. patient is unequivocally false.”

When Mrs. Hira arrived at the University of Chicago, it was clear that she was a V.I.P., according to two people involved in her care who requested anonymity because they were not authorized to discuss patients.

Mrs. Hira, then 55, was married to Ryuko Hira, the owner of the HMI Hotel Group, one of Japan’s largest hotel chains. She and her husband also had something in common with the hospital’s top heart surgeon, Dr. Valluvan Jeevanandam. They were all followers of the Indian spiritual figure Sathya Sai Baba.

She had faced long odds of getting a new heart in Japan, where only about 10 percent of residents are registered donors and the wait for a transplant can stretch for years.

On Sept. 30, 2021, the University of Chicago added Mrs. Hira to the registry and obtained an exception that increased her priority level, records show.

The same day, a 19-year-old woman died in the Midwest. The transplant system’s algorithm identified more than 400 potential matches for her heart — including Mrs. Hira.

Patients often wait months to gain priority and for the right organ to become available. But with the exception, Mrs. Hira was No. 7 on the list. Doctors for the patients above her determined the organ wasn’t right for them, and then the University of Chicago accepted. Dr. Jeevanandam performed the transplant on Oct. 3.

Mrs. Hira got her transplant faster than 91 percent of heart recipients that year, data shows.

She was a “self-pay” patient, meaning she didn’t use insurance, according to records reviewed by The Times. Such international patients often pay the full billed amount, which averages about $1.9 million for a heart transplant, according to recent research.

The month after the transplant, the Sai Hira India Foundation, a charity founded by Mr. Hira, donated money to the Sai Spiritual Foundation, a Chicago nonprofit that promotes the teachings of Sai Baba and performs community service. Tax filings show that the Chicago nonprofit’s principal officer was Sheela Jeevanandam, the surgeon’s wife. Public records do not disclose the amount of the donation; that year, the group reported raising about $1 million more than any year before or since.

The University of Chicago declined to discuss Mrs. Hira’s case, citing patient privacy laws. A representative for Mr. Hira declined to comment, and Dr. Jeevanandam did not respond to requests for comment.

Dr. Gabriel Danovitch, a longtime leader of the U.S. transplant system and a co-author of the 2012 policy that allowed for more international patients, said that transplant tourism was still very uncommon in America. But he said he was greatly disturbed to hear that an overseas patient had so quickly received an organ and made a donation. “Truthfully, it makes me sick,” he said.

About three weeks after her transplant, Mrs. Hira had recovered enough to leave the hospital. She told her caregivers there that before returning to Japan, she wanted to give them something to remember her. They each received a jacket embroidered with their name, the date of Mrs. Hira’s transplant and the lotus flower symbol of Sai Baba.

Brian M. Rosenthal

https://www.nytimes.com/2025/12/16/us/organ-transplants-international-patients.html?unlocked_article_code=1.9E8.dO6a.LDE8Ox-4zVKp&smid=url-share




Friday, December 12, 2025

Levacetylleucine treatment of Niemann-Pick disease type C

N-acetyl-L-leucine (Levacetylleucine) normalizes Transcription Factor EB (TFEB) activity by stereospecific bidirectional modulation
Lianne C. Davis, Rebecca Braine, Grant C. Churchill, Mallory Factor, Taylor Fields, Marc Patterson, Frances Platt, Michael Strupp, Antony Galione. bioRxiv 2025.11.30.691375; doi: https://doi.org/10.64898/2025.11.30.691375

This article is a preprint and has not been certified by peer review

Abstract

Levacetylleucine (AqneursaTM), a chemically modified amino acid, is the only US Food and Drug Administration-approved monotherapy for the treatment of Niemann-Pick disease type C (NPC) (Beninger, 2024; Mullard, 2024; van Gool et al., 2025). This acetylated derivative of L-leucine functions as a pro-drug, with the acetyl group rendering it a substrate for the monocarboxylate transporter (MCT) family of transporters to allow appreciable penetration of the blood-brain barrier and its efficient uptake into cells (Churchill et al., 2021). Inside cells, levacetylleucine undergoes metabolism catalysed by acylases, and the resultant high quantities of L-leucine enter metabolic pathways which enhance mitochondrial bioenergetics and, as previously demonstrated, indirectly ameliorate lysosomal function (Kaya et al., 2020). Here, we show a novel aspect of levacetylleucine’s mechanism of action, demonstrating a direct effect on lysosomal function through its rapid modulation of the translocation of the transcription factor TFEB, a master regulator of lysosomal biogenic and autophagic genes (Napolitano and Ballabio, 2016), from cytoplasm to nucleus. Uniquely, we have demonstrated a biphasic action whereby levacetylleucine normalizes TFEB activity, consistent with levacetylleucine’s previously shown ability to regulate cellular homeostasis: in wild-type HeLa cells, levacetylleucine enhances and activates the translocation of TFEB to the nucleus. In contrast, in cellular models of NPC type 1 disease, where TFEB is already over-expressed in the nucleus (as the cell attempts to compensate for the primary defect by activating TFEB as a natural cellular response to the lysosomal substrate accumulation and associated cellular stress), treatment with levacetylleucine down-regulates and restores the distribution of TFEB to a more normalized cytoplasmic: nuclear ratio. Importantly, both effects of levacetylleucine occur at concentrations consistent with plasma concentrations in therapeutic dosing (Churchill et al., 2020). The effects were also confirmed to be stereospecific to the L-enantiomer, as neither the D-enantiomer (N-acetyl-D-leucine) or racemate (N-acetyl-DL-leucine) had any effect, The presence of the D-enantiomer in the racemic mixture inhibited the ability of levacetylleucine to promote TFEB bidirectional translocation, consistent with previous studies, which have established antagonism of N-acetyl-L-leucine by N-acetyl-D-leucine in the racemic mixture (rendering the racemic mixture without effect). This bidirectional mechanism of action of levacetylleucine to impact lysosomal function directly and normalize, either by activating basal TFEB signalling or reducing aberrant TFEB function in NPC1 knockout cells, thereby modulating lysosomal and autophagic functions, lends itself to the treatment of a broad range of neurological and neurodevelopment disorders.

Thursday, December 11, 2025

Physicians receiving payments are more likely to prescribe the company’s drugs

Sayed A, Gupta R, Ramachandran R, et al. Industry payments to US neurologists related to multiple sclerosis drugs and prescribing (2015–2019): a retrospective cohort study. BMJ Open 2025;15:e095952. doi: 10.1136/bmjopen-2024-095952

Abstract

Objectives To measure the prevalence and magnitude of industry payments to neurologists prescribing multiple sclerosis (MS) drugs and determine whether payments are associated with prescribing.

Design Retrospective observational study.

Setting Data on neurologists prescribing MS drugs from 2015 to 2019 in the Medicare Part D database linked to the Centers for Medicare & Medicaid Services Open Payments database.

Participants 7401 neurologists prescribing MS drugs from 2015 to 2019 to Medicare beneficiaries.

Main outcome measures The primary outcome was the proportion of physicians’ annual prescriptions manufactured by a given company. Generalised linear mixed models were used to evaluate associations between the presence and magnitude of payments and prescribing. The association between prescription volume and the likelihood of receiving payments as well as the value of payments was also assessed.

Results Among 7401 neurologists, 5809 (78.5%) received payments totalling US$163.6 million between 2015 and 2019. While the median amount per physician was US$779 (IQR, US$188–US$2587), US$155.7 million (95.2%) accrued to the top 10% of payment recipients. Higher prescription volumes were associated with a higher likelihood of receiving any payment type, particularly for consulting services, non-consulting services and travel/lodging (p<0.001). Among payment recipients, the amount received was positively associated with prescription volume (p<0.001). Receipt of payments was associated with greater likelihood of prescribing the company’s drugs compared with those who received no payments from that company (OR 1.13 (95% CI 1.11 to 1.15)), with the largest association for non-consulting services, such as being a speaker at an event (OR 1.53 (95% CI 1.44 to 1.62)). Larger payments were associated with a greater likelihood of prescribing (OR 1.10, 1.26, 1.29 and 1.50 for US$50, US$500, US$1000 and US$5000, respectively), as were longer durations of payments (OR 1.12 for single year to 1.78 for 5 consecutive years) and more recent payments (OR 1.03 for payments made 4 years prior to 1.34 for payments made in the same year).

Conclusions Nearly 80% of neurologists prescribing MS drugs received at least one industry payment, with higher volume prescribers being more likely to receive payments. Physicians receiving payments were more likely to prescribe the company’s drugs, with a stronger association for payments that were larger, sustained and recent.
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A ‘Humbling’ Study

Two MS neurologists without ties to the research—Elizabeth Silbermann, MD, an assistant professor of neurology at Oregon Health & Sciences University (OHSU), and Mitchell Wallin, MD, MPH, FAAN, associate professor of neurology at the University of Maryland School of Medicine and director of the VA MS Center of Excellence—said Dr. Ross and his team made a good analysis of the data available to them.

“It's a good population-based sample of the United States with high numbers,” said Dr. Wallin, who thought it was wise that the researchers stopped the analysis at the beginning of the pandemic. He added that he was impressed that the study showed a dose-response relationship between payments and prescribing. “I think that it's a humbling article.”

Dr. Silberman called it a broad study that “captured data in the best way you can in the United States.” She said the high percentage of neurologists who accepted pharma money had a “bit of shock value.”
The data problem, which the researchers acknowledged, is that the study was limited to prescribing information from Medicare Part D. MS tends to be diagnosed among much younger people, so the study may have missed those just starting medications as well as some younger people with aggressive disease.

In addition, Dr. Wallin said that many MS experts at academic medical centers are now trying to wean older, stable patients from their medications. And newer infusion drugs are paid through Medicare Part B, so Part D data does not capture them.

Dr. Ross acknowledged that the data his team used skewed toward older patients and young ones with serious disabilities. However, the MS specialists included in the study also treat younger patients, and he thinks their prescribing habits likely transcended age. It is possible that industry influence on prescribing behavior is even more pronounced among younger patients because they change drugs more frequently, he said.

Dr. Wallin pointed out that insurance companies often resist paying for newer, more expensive drugs, which may mean that doctors end up prescribing medications that were not their first or even second choice.

Earlier this year, Dr. Silbermann's colleagues at OHSU published a study on uptake of generic glatiramer acetate and industry payments that found an association between the payments and prescriptions for the branded drug. Although generics became available in 2015, more than half of prescriptions in 2021 were still for the branded formulation. Neurologists who received industry payments were more likely to prescribe the branded drug, the study found.
One confounding factor, Dr. Silbermann said, is that drug companies often provide financial assistance for patients who take the branded drug.

Why Does This Problem Persist?

Asked why doctors continue to accept payments from drug companies when multiple studies have shown that it affects prescribing behavior, Dr. Ross said he has heard from other doctors that they don't think payments can influence them. Also, it's part of medical culture. Doctors think, “Everyone's doing it, so why shouldn't I?” he said.

Dr. Wallin, who does not accept direct payments from pharma, agreed that most physicians “believe that pharmaceutical reps don't really affect their behavior,” but “people are avaricious. That's the short answer.”

Unfortunately, he said, medical groups depend on industry to support their large, educational meetings, and he and Dr. Silbermann agreed that talking to sales reps can be valuable. She occasionally attends educational events sponsored by a drug company but doesn't eat or drink, except for a cappuccino she might accept from a drug company at a big meeting. Dr. Silbermann said she understands why hungry, time-pressed convention-goers might accept some food, though.

The big issue, she said, is that doctors need information about drugs, and it's hard to get it from primary sources. Keeping up to date on more than 20 medications might require reading 50 papers a year.
“Pharma picked up on a gap in the field, which is that physicians need to receive high-quality information in order to inform their practices,” she said. “It's really hard to receive high-quality information for free these days. Conferences are incredibly expensive.”

While many payments are small, they add up over time. Dr. Ross believes companies wouldn't spend the money if they didn't think the tactic was working.

“These companies aren't making these payments for nothing,” he said.

He had hoped that federal rules requiring disclosure of payments would make physicians stop taking them. At first, doctors worried about having their names and payment amounts made public. The information has been used for research, but fears of widespread embarrassment have not materialized.
“For the most part, it's kind of out of sight, out of mind,” said Dr. Ross, adding that half measures are unlikely to make a dent in the problem. “I think companies continue to engage in these practices because physicians engage in these practices. The way you would fix it would be to declare all payments to be in violation of kickback statutes and say they're illegal.” Current government guidance, he added, is “fuzzy.”

While Dr. Wallin thinks it makes sense to limit contact between drug company representatives and doctors in clinical practice, he wouldn't want to cut off all communication between drug makers and physicians. It's helpful to discuss side effects with company employees, he said. A middle ground might be an educational nonprofit that could accept pharma money but would not tie specific talks to an individual company, Dr. Wallin said.

National conferences, along with podcasts and journal groups from the AAN, can help physicians stay informed, Dr. Silbermann noted. A good starting point for further improvements, she said, would be for neurologists to raise awareness that an educational gap needs to be filled in a way that's less likely to influence physician decision-making.

Disclosures:
Dr. Ross was an expert witness at the request of Relator's attorneys, the Greene Law Firm, in a qui tam suit alleging violations of the False Claims Act and Anti-Kickback Statute against Biogen Inc. that was settled September 2022. He currently receives research support through Yale University from Johnson & Johnson to develop methods of clinical trial data sharing, from the Food and Drug Administration for the Yale-Mayo Clinic Center for Excellence in Regulatory Science and Innovation (CERSI) program (U01FD005938), from the Greenwall Foundation, and from Arnold Ventures. Dr. Wallin is a principal investigator on a Sanofi-sponsored randomized controlled drug trial and received travel funding as a speaker from CMSC.

https://neurologytoday.aan.com/doi/10.1097/01.wnt.0001173580.24350.4e

Wednesday, December 10, 2025

ANKLE2 autosomal recessive congenital microcephaly

Inspired by sibling patients

Fishburn AT, Florio CJ, Lopez NJ, Link NL, Shah PS. Molecular functions of ANKLE2 and its implications in human disease. Dis Model Mech. 2024 Apr 1;17(4):dmm050554. doi: 10.1242/dmm.050554. Epub 2024 May 1. PMID: 38691001; PMCID: PMC11103583.

Abstract

Ankyrin repeat and LEM domain-containing 2 (ANKLE2) is a scaffolding protein with established roles in cell division and development, the dysfunction of which is increasingly implicated in human disease. ANKLE2 regulates nuclear envelope disassembly at the onset of mitosis and its reassembly after chromosome segregation. ANKLE2 dysfunction is associated with abnormal nuclear morphology and cell division. It regulates the nuclear envelope by mediating protein-protein interactions with barrier to autointegration factor (BANF1; also known as BAF) and with the kinase and phosphatase that modulate the phosphorylation state of BAF. In brain development, ANKLE2 is crucial for proper asymmetric division of neural progenitor cells. In humans, pathogenic loss-of-function mutations in ANKLE2 are associated with primary congenital microcephaly, a condition in which the brain is not properly developed at birth. ANKLE2 is also linked to other disease pathologies, including congenital Zika syndrome, cancer and tauopathy. Here, we review the molecular roles of ANKLE2 and the recent literature on human diseases caused by its dysfunction.

Fishburn AT, Florio CJ, Klaessens TN, Prince B, Adia NAB, Lopez NJ, Beesabathuni NS, Becker SS, Cherkashchenko L, Haggard Arcé ST, Hoang V, Shiu TN, Richardson RB, Evans MJ, Rückert C, Shah PS. Microcephaly protein ANKLE2 promotes Zika virus replication. mBio. 2025 Feb 5;16(2):e0268324. doi: 10.1128/mbio.02683-24. Epub 2025 Jan 13. PMID: 39804047; PMCID: PMC11796389.

Abstract

Orthoflaviviruses are positive-sense single-stranded RNA viruses that hijack host proteins to promote their own replication. Zika virus (ZIKV) is infamous among orthoflaviviruses for its association with severe congenital birth defects, notably microcephaly. We previously mapped ZIKV-host protein interactions and identified the interaction between ZIKV non-structural protein 4A (NS4A) and host microcephaly protein ankyrin repeat and LEM domain-containing 2 (ANKLE2). Using a fruit fly model, we showed that NS4A induced microcephaly in an ANKLE2-dependent manner. Here, we explore the role of ANKLE2 in ZIKV replication to understand the biological significance of the interaction from a viral perspective. We observe that ANKLE2 localization is drastically shifted to sites of NS4A accumulation during infection and that knockout of ANKLE2 reduces ZIKV replication in multiple human cell lines. This decrease in virus replication is coupled with a moderate increase in innate immune activation. Using microscopy, we observe dysregulated formation of virus-induced endoplasmic reticulum rearrangements in ANKLE2 knockout cells. Knockdown of the ANKLE2 ortholog in Aedes aegypti cells also decreases virus replication, suggesting ANKLE2 is a beneficial replication factor across hosts. Finally, we show that NS4A from four other orthoflaviviruses physically interacts with ANKLE2 and is also beneficial to their replication. Thus, ANKLE2 likely promotes orthoflavivirus replication by regulating membrane rearrangements that serve to accelerate viral genome replication and protect viral dsRNA from immune detection. Taken together with our previous results, our findings indicate that ZIKV and other orthoflaviviruses hijack ANKLE2 for a conserved role in replication, and this drives unique pathogenesis for ZIKV since ANKLE2 has essential roles in developing tissues.IMPORTANCEZIKV is a major concern due to its association with birth defects, including microcephaly. We previously identified a physical interaction between ZIKV NS4A and host microcephaly protein ANKLE2. Mutations in ANKLE2 cause congenital microcephaly, and NS4A induces microcephaly in an ANKLE2-dependent manner. Here, we establish the role of ANKLE2 in ZIKV replication. Depletion of ANKLE2 from cells significantly reduces ZIKV replication and disrupts virus-induced membrane rearrangements. ANKLE2's ability to promote ZIKV replication is conserved in mosquito cells and for other related mosquito-borne orthoflaviviruses. Our data point to an overall model in which ANKLE2 regulates virus-induced membrane rearrangements to accelerate orthoflavivirus replication and avoid immune detection. However, ANKLE2's unique role in ZIKV NS4A-induced microcephaly is a consequence of ZIKV infection of important developing tissues in which ANKLE2 has essential roles.

Shaheen R, Maddirevula S, Ewida N, Alsahli S, Abdel-Salam GMH, Zaki MS, Tala SA, Alhashem A, Softah A, Al-Owain M, Alazami AM, Abadel B, Patel N, Al-Sheddi T, Alomar R, Alobeid E, Ibrahim N, Hashem M, Abdulwahab F, Hamad M, Tabarki B, Alwadei AH, Alhazzani F, Bashiri FA, Kentab A, Şahintürk S, Sherr E, Fregeau B, Sogati S, Alshahwan SAM, Alkhalifi S, Alhumaidi Z, Temtamy S, Aglan M, Otaify G, Girisha KM, Tulbah M, Seidahmed MZ, Salih MA, Abouelhoda M, Momin AA, Saffar MA, Partlow JN, Arold ST, Faqeih E, Walsh C, Alkuraya FS. Genomic and phenotypic delineation of congenital microcephaly. Genet Med. 2019 Mar;21(3):545-552. doi: 10.1038/s41436-018-0140-3. Epub 2018 Sep 14. PMID: 30214071; PMCID: PMC6986385.

Abstract

Purpose: Congenital microcephaly (CM) is an important birth defect with long term neurological sequelae. We aimed to perform detailed phenotypic and genomic analysis of patients with Mendelian forms of CM.

Methods: Clinical phenotyping, targeted or exome sequencing, and autozygome analysis.

Results: We describe 150 patients (104 families) with 56 Mendelian forms of CM. Our data show little overlap with the genetic causes of postnatal microcephaly. We also show that a broad definition of primary microcephaly -as an autosomal recessive form of nonsyndromic CM with severe postnatal deceleration of occipitofrontal circumference-is highly sensitive but has a limited specificity. In addition, we expand the overlap between primary microcephaly and microcephalic primordial dwarfism both clinically (short stature in >52% of patients with primary microcephaly) and molecularly (e.g., we report the first instance of CEP135-related microcephalic primordial dwarfism). We expand the allelic and locus heterogeneity of CM by reporting 37 novel likely disease-causing variants in 27 disease genes, confirming the candidacy of ANKLE2, YARS, FRMD4A, and THG1L, and proposing the candidacy of BPTF, MAP1B, CCNH, and PPFIBP1.

Conclusion: Our study refines the phenotype of CM, expands its genetics heterogeneity, and informs the workup of children born with this developmental brain defect.

Naveed M, Kazmi SK, Amin M, Asif Z, Islam U, Shahid K, Tehreem S. Comprehensive review on the molecular genetics of autosomal recessive primary microcephaly (MCPH). Genet Res (Camb). 2018 Aug 8;100:e7. doi: 10.1017/S0016672318000046. PMID: 30086807; PMCID: PMC6865151.

Abstract

Primary microcephaly (MCPH) is an autosomal recessive sporadic neurodevelopmental ailment with a trivial head size characteristic that is below 3-4 standard deviations. MCPH is the smaller upshot of an architecturally normal brain; a significant decrease in size is seen in the cerebral cortex. At birth MCPH presents with non-progressive mental retardation, while secondary microcephaly (onset after birth) presents with and without other syndromic features. MCPH is a neurogenic mitotic syndrome nevertheless pretentious patients demonstrate normal neuronal migration, neuronal apoptosis and neural function. Eighteen MCPH loci (MCPH1-MCPH18) have been mapped to date from various populations around the world and contain the following genes: Microcephalin, WDR62, CDK5RAP2, CASC5, ASPM, CENPJ, STIL, CEP135, CEP152, ZNF335, PHC1, CDK6, CENPE, SASS6, MFSD2A, ANKLE2, CIT and WDFY3, clarifying our understanding about the molecular basis of microcephaly genetic disorder. It has previously been reported that phenotype disease is caused by MCB gene mutations and the causes of this phenotype are disarrangement of positions and organization of chromosomes during the cell cycle as a result of mutated DNA, centriole duplication, neurogenesis, neuronal migration, microtubule dynamics, transcriptional control and the cell cycle checkpoint having some invisible centrosomal process that can manage the number of neurons that are produced by neuronal precursor cells. Furthermore, researchers inform us about the clinical management of families that are suffering from MCPH. Establishment of both molecular understanding and genetic advocating may help to decrease the rate of this ailment. This current review study examines newly identified genes along with previously identified genes involved in autosomal recessive MCPH.