Skip to main content

Natural history, outcome measures and trial readiness in LAMA2-related muscular dystrophy and SELENON-related myopathy in children and adults: protocol of the LAST STRONG study



SELENON (SEPN1)-related myopathy (SELENON-RM) is a rare congenital myopathy characterized by slowly progressive proximal muscle weakness, early onset spine rigidity and respiratory insufficiency. A muscular dystrophy caused by mutations in the LAMA2 gene (LAMA2-related muscular dystrophy, LAMA2-MD) has a similar clinical phenotype, with either a severe, early-onset due to complete Laminin subunit α2 deficiency (merosin-deficient congenital muscular dystrophy type 1A (MDC1A)), or a mild, childhood- or adult-onset due to partial Laminin subunit α2 deficiency. For both muscle diseases, no curative treatment options exist, yet promising preclinical studies are ongoing. Currently, there is a paucity on natural history data and appropriate clinical and functional outcome measures are needed to reach trial readiness.


LAST STRONG is a natural history study in Dutch-speaking patients of all ages diagnosed with SELENON-RM or LAMA2-MD, starting August 2020. Patients have four visits at our hospital over a period of 1.5 year. At all visits, they undergo standardized neurological examination, hand-held dynamometry (age ≥ 5 years), functional measurements, questionnaires (patient report and/or parent proxy; age ≥ 2 years), muscle ultrasound including diaphragm, pulmonary function tests (spirometry, maximal inspiratory and expiratory pressure, sniff nasal inspiratory pressure; age ≥ 5 years), and accelerometry for 8 days (age ≥ 2 years); at visit one and three, they undergo cardiac evaluation (electrocardiogram, echocardiography; age ≥ 2 years), spine X-ray (age ≥ 2 years), dual-energy X-ray absorptiometry (DEXA-)scan (age ≥ 2 years) and full body magnetic resonance imaging (MRI) (age ≥ 10 years). All examinations are adapted to the patient’s age and functional abilities. Correlation between key parameters within and between subsequent visits will be assessed.


Our study will describe the natural history of patients diagnosed with SELENON-RM or LAMA2-MD, enabling us to select relevant clinical and functional outcome measures for reaching clinical trial-readiness. Moreover, our detailed description (deep phenotyping) of the clinical features will optimize clinical management and will establish a well-characterized baseline cohort for prospective follow-up.


Our natural history study is an essential step for reaching trial readiness in SELENON-RM and LAMA2-MD.

Trial registration

This study has been approved by medical ethical reviewing committee Region Arnhem-Nijmegen (NL64269.091.17, 2017–3911) and is registered at (NCT04478981).

Peer Review reports


Selenoprotein N-related congenital myopathy (SEPN1- or SELENON-RM) is a rare congenital myopathy with an estimated prevalence of 0.5 in 1000,000 [1]. Core features include slowly progressive axial muscle weakness, early-onset rigidity of the spine, scoliosis and respiratory insufficiency. Delayed motor development is the most common presenting sign. Muscle biopsies show multi-minicores as the most common lesion, often associated with mild dystrophic features [2]. Laminin α2-related muscular dystrophy (LAMA2-MD) has a similar clinical phenotype, with an estimated prevalence of 4 in 500,000 [3]. It has a heterogeneous disease spectrum ranging from a severe, early-onset congenital muscular dystrophy (complete Laminin subunit α2 deficiency, also called merosin-deficient congenital muscular dystrophy type 1A (MDC1A)) to a mild, childhood- or adult-onset limb-girdle type muscular dystrophy (partial Laminin subunit α2 deficiency). Additionally, patients may suffer from epileptic seizures and may show characteristic diffuse brain white matter lesions on magnetic resonance imaging (MRI) [4]. The clinical diagnosis of SELENON-RM and LAMA2-MD is confirmed by recessive pathogenic variants in the SELENON gene (OMIM-number 606210) or LAMA2 gene (OMIM-number 156225), respectively [5,6,7,8,9]. Currently, no curative treatment options exist for neither SELENON-RM nor LAMA2-MD. Optimal pulmonary, cardiac, nutrition and orthopedic management, in combination with supportive care from rehabilitation and allied health care, is essential for preventing or treating severe complications [10, 11]. Further, promising new therapies are currently being developed [12,13,14,15,16,17,18,19,20,21].

Selenoprotein N is an endoplasmic reticulum (ER) calcium sensor that responds to diminished luminal calcium levels by refilling the ER calcium stores [22]. SELENON-RM has striking similarities at the cellular level with classical mitochondrial diseases. This has led to the hypothesis that sonlicromanol, a new clinical stage chemical entity with a dual activity as antioxidant and redox modulator developed for mitochondrial oxidative phosphorylation disturbances, is also beneficial for patients with SELENON-RM [12, 13]. Interestingly, the first results of experiments in an animal model (SELENON knock-out zebrafish) showed improved muscular function (unpublished data). Further, other antioxidants are also hypothesized to be beneficial in the treatment of patients with SELENON-RM [13,14,15]. In general, the recognition of the interplay between mitochondrial bioenergetics and endoplasmic reticulum paves the way to the identification of potential treatment options [16, 17]. Laminin subunit α2 is an extracellular matrix protein that links with dystrophin on the inner side of the muscle membrane. This linkage is of high importance for normal skeletal muscle function as it stabilizes the sarcolemma and protects the muscle fibers from contraction-induced damage [23, 24]. Further, a metabolic impairment, with reduced mitochondrial respiration and enhanced glycolysis, was observed in human Laminin subunit α2 deficient muscle cells [25]. For LAMA2-MD, preclinical studies on the use of linker proteins, on exogenous administration of Laminin-111, on upregulation of LAMA1, on genome editing technology and on the use of antioxidant molecules are being performed in animal models [18,19,20,21, 26, 27]. Moreover, different research groups are working on revealing the precise pathophysiology, which could eventually help to design new treatment strategies for SELENON-RM [14, 28,29,30,31] or LAMA2-MD [32,33,34].

In order to pave the way towards clinical trials, it is essential to identify and characterize the patients clinically and genetically, and to select clinical and functional outcome measures that correlate with muscle function and that are sensitive to change over time. These outcome measures can be used to determine the effectivity of possible treatment options in clinical trials. Three large clinical studies have recently been performed, one in SELENON-RM and two in LAMA2-MD. We discuss the main findings below.

In SELENON-RM patients, a retrospective clinical, histologic and genetic analysis of 132 pediatric and adult patients (age range at last examination 2 to 58 years) was performed by Villar-Quiles et al. [2]. The main prognostic determinants for disease severity included scoliosis and respiratory management, body mass abnormalities and the specific SELENON mutation found in the patient. The latter indicates a genotype-phenotype association between bi-allelic null mutations and more severe disease. This study reports the largest SELENON-RM series and the first one including pediatric, adolescent and adult patients followed-up for several decades. Limitations of this study are its retrospective design, and the absence of functional measurements and convenient muscle visualizing techniques (i.e. muscle ultrasound or MRI) performed in a standardized manner.

In LAMA2-MD patients, a 5-year prospective natural history study that included 24 patients (age range 4 to 22 years) was performed by Jain et al. [35, 36]. The MFM-32 was found to be sensitive to change in ambulatory and non-ambulatory patients with LAMA2-MD. In non-ambulatory patients, they found a yearly decline in knee flexion strength and passive range of motion (PROM) of left elbow extension. Limitations of this study included the small subset of examinations performed and the absence of convenient muscle visualizing techniques. Further, patients were selected based on a convenience sample, possibly leading to a selection bias. Moreover, the age range limits the availability on natural history data in the very young children (< 4 years) and in older adult patients (> 22 years). Recently, Zambon et al. published a retrospective longitudinal study on 46 patients with LAMA2-MD of whom 42 patients had a complete and 4 patients had a partial Laminin subunit α2 deficiency (age range at last examination 12 to 22 years) [37]. They found a linear decrease in passive range of motion of left elbow extension and a linear decline in percentage predicted forced vital capacity. The intrinsic limitations included the retrospective nature of data collection, the limited age range and the inconsistencies in the use of functional scales throughout follow-up (i.e. frequency of examinations, indication for ancillary examinations etcetera).

In short, a prospective natural history study in an unselected group of patients including a plethora of clinical and functional outcome measures is lacking in both SELENON-RM and LAMA2-MD. Due to the promising ongoing preclinical studies, there is a high need to obtain natural history data in order to reach trial readiness for both muscle diseases. The similarities in the clinical phenotype of both muscle diseases allows us to combine both studies in one study protocol.


The primary objectives of this study in order to reach trials readiness, are:

  1. 1.

    to assess 1.5-year natural history in patients with SELENON-RM or LAMA2-MD;

  2. 2.

    to select relevant and sensitive clinical and functional outcome measures.

The secondary objectives of this study are:

  1. 3.

    to provide prevalence estimations of SELENON-RM and LAMA2-MD in the Netherlands and Flanders (Dutch-speaking part of Belgium);

  2. 4.

    to establish a well-characterized baseline cohort of patients with SELENON-RM or LAMA2-MD for prospective follow-up and recruitment for future clinical trials;

  3. 5.

    to assess the clinical features to optimize clinical management for patients with SELENON-RM or LAMA2-MD.

Methods / design

Study design

Our study on LAMA2-MD and SELENON-RM To Study Trial Readiness, Outcome measures and Natural history (LAST STRONG) is a prospective, single-center, observational study with repeated measurements performed at the Department of Neurology and Pediatric Neurology within the neuromuscular center of the Radboud university medical center, The Netherlands. Our center is a tertiary referral center for neuromuscular diseases. Participation in the study will not affect the usual care provided by the patient’s own medical team. Patients are invited to visit our hospital four times over a period of 1.5 year, with an interval of six months. During these visits, a predefined subset of investigations will be performed (See Table 1). Further, medical records from routine clinical care will be requested. Our study has been approved by the medical ethical reviewing committee Region Arnhem – Nijmegen (NL64269.091.17; 2017–3911) and is registered at (NCT04478981).

Table 1 Examinations performed in LAST STRONG study

Study population

Based on the prevalence reported in previous studies on SELENON-RM and LAMA2-MD, we expect to identify between 20-30 patients in each disease group. Based on our experience in rare diseases, we estimate that around 50% of the patients will participate in the study. We therefore aim to include 10-15 participants in each disease group. Inclusion criteria include a genetic confirmation of SELENON-RM or LAMA2-MD by two recessive pathologic mutations in the SELENON or LAMA2 gene, respectively, or typical clinical and histological alterations combined with genetic confirmation in a first degree relative. Additionally, patients must be willing and able to complete (part of) the measurement protocol in the Radboud university medical center. If patients do not wish or are not able to visit our center, they are offered to participate in this study by sharing medical records, completing questionnaires, and undergoing a medical history and physical examination through home visits, video and/or telephone interview. Exclusion criteria are an insufficient understanding of the Dutch language and the unwillingness of the patient or his/her legal representatives to provide written informed consent for participation in our study.


Participants will be recruited non-selectively and consecutively in the periods from August 2020 to August 2021 (See Fig. 1). In order to estimate the prevalence and to provide data on the complete spectrum of patients with SELENON-RM or LAMA2-MD, we aim to reach all patients in The Netherlands and the Dutch-speaking part of Belgium (Flanders) in our study. All Dutch and Dutch-speaking Belgian (pediatric) neurologists, rehabilitation specialists and clinical geneticists are personally asked about potential participants. Additionally, all patients known at our own neuromuscular center will be personally informed. Moreover, we will directly recruit participants through promotion of our study on patient information days, social media and through patient organizations.

Fig. 1
figure 1

Flow chart of recruitment and inclusion of SELENON-RM or LAMA2-MD patients


Date of birth, sex, weight (kg), height (m), comorbidity and medication will be recorded.


Upon inclusion in our study, all participants (or a first degree relative) have undergone genetic examination as part of regular diagnostic work-up. The genetics reports, including information on the specific genetic alterations, will be requested. Regarding LAMA2-MD, we expect to mostly include patients harboring the Dutch founder mutation in the LAMA2 gene (c.5562 + 5G > C). All genetic laboratories in The Netherlands will be contacted to inventory the number of patients diagnosed with SELENON-RM or LAMA2-MD in order to contrast this number against the number of participants in our natural history study.

Muscle biopsy

Pathological records and stained slides will be requested from participants diagnosed with LAMA2-MD in whom muscle biopsy material was previously taken as part of regular diagnostic work-up. Hereby we aim to classify LAMA2-MD patients as either suffering from a complete or a partial Laminin subunit α2 deficiency.

Neurological examination and functional measurements

All patients undergo a standard neurological examination by one assessor (KB). Additionally, muscle strength, facial muscle weakness, reflexes, muscle tone, and dysmorphic features are assessed by two independent assessors (KB and CE or NV). Muscle strength (MRC grading scale) will be assessed of the following muscles: neck flexor, neck extensor, sternocleidomastoid, trapezius, deltoid, biceps brachii, triceps brachii, wrist extensor, wrist flexor, finger extensor, finger flexor, finger spreader, iliopsoas, gluteus maximus, quadriceps, hamstrings, foot dorsiflexor, foot plantarflexor, extensor hallucis longus and toe flexor muscles. Further, muscle strength of the following muscles will be measured using a hand-held dynamometer (Citec, CT3002) [38,39,40,41]:

  1. 1.

    Neck flexors and neck extensors: sitting upright; head up at 90° from horizontal

  2. 2.

    Elbow flexors and extensors: supine; shoulder adducted, elbow 90° flexed, forearm supinated

  3. 3.

    Knee extensors: sitting upright; knee 90° flexed

  4. 4.

    Foot plantar- and dorsiflexors: supine; foot 90° dorsiflexed

  5. 5.

    Pinch grip: sitting upright; shoulder adducted, elbow 90° flexed, forearm pronated

Additionally, the passive range of motion (PROM) of the elbow, wrist, hip, knee and ankle joints is assessed by a goniometer [42]. Functional measurements include:

  1. 1.

    The Children’s Hospital of Philadelphia Infant Test of Neuromuscular Disorders (CHOP INTEND) (age < 2 years) [43, 44].

    1. a.

      CHOP INTEND has been shown to be valid for the assessment of motor skills of children below 2 years of age.

  2. 2.

    Hammersmith Infant Neurological Examinations (HINE) (age < 2 years) [45].

    1. a.

      HINE is designed to be a simple and scorable method for evaluating infants from 2 months to 2 years of age. It includes three sections that assess different aspects of neurologic function, including neurological examination, developmental milestones and behavioral assessment.

  3. 3.

    Motor Function Measure – 20/32 (MFM-20/32) (age ≥ 2 years) [46, 47].

    1. a.

      Motor function in patients with neuromuscular diseases can be measured with the MFM. The MFM is a scale which consists of 20 or 32 items in three dimensions: D1: standing position and transfers, D2: axial and proximal motor function, D3: distal motor function. MFM-32 is used in adults and in children of 7 years and older. Children with the age of 2 to 7 years will undergo MFM-20.

  4. 4.

    Hammersmith Functional Motor Scale (HFMS) (age ≥ 2 years; non-ambulant participants only) [48, 49].

    1. a.

      The HFMS was originally developed to assess the physical abilities of children with non-ambulant spinal muscular atrophy. It consists of 20 items that were considered as important to measure the physical functioning of those patients.

  5. 5.

    Pediatric Balance Scale (PBS) (pediatric patients aged 2–15 years; ambulant participants only) [50].

    1. a.

      The PBS is a modified version of the Berg Balance Scale that is used to assess functional balance skills in school-aged children with mild to moderate motor impairments.

  6. 6.

    Mini Balance Evaluation Systems Test (miniBEST) (age ≥ 16 years; ambulant participants only) [51].

    1. a.

      The miniBEST evaluates balance control by scoring of exercises that belong to one of the following categories: anticipatory postural changes, reactive postural control, sensory orientation and walking.

  7. 7.

    Graded and timed function tests (age ≥ 2 years: 30 s sit to stand, climb 4 stairs, rise from the floor, timed up and go (TUG); age ≥ 5 years: 6-Minute Walk test, 10-Meter Walk test; ambulant participants only) [52,53,54].

  8. 8.

    Functional Ambulation Classification (FAC) (age ≥ 5 years) [55].

    1. a.

      The FAC assesses functional ambulation in patients.

  9. 9.

    Brooke and Vignos scale (age ≥ 2 years) [56,57,58].

    1. a.

      The Brooke and Vignos scales provide ordinal-level data to assess the upper and lower extremity functions, respectively.

Neurological examination and functional measurements will be performed at all four visits.


Each visit, patients and/or their parent(s) will be asked to complete age-adapted questionnaires on quality of life, pain, fatigue, and activities and participation,. These questionnaires include:

  1. 1.

    Pediatric Quality of Life Inventory (PedsQL) (Generic Core Scale, Neuromuscular Module, Multidimensional Fatigue Scale; Child Self-report and/or parent proxy) (pediatric patients 2–17 years) [59,60,61].

    1. a.

      The PedsQL Generic Core Scale consists of 23 questions in four domains: Physical, Emotional, Social, and School Functioning. It has been translated and subsequently validated into many languages, including Dutch.

    2. b.

      The PedsQL Neuromuscular Module consists of 25 questions in three domains: Neuromuscular disease, Communication and Family resources

    3. c.

      The PedsQL Multidimensional Fatigue Scale assesses subjective fatigue in three domains, namely General Fatigue Scale, Sleep/Rest Fatigue Scale, and Cognitive Fatigue Scale.

  2. 2.

    Research and Development-36 (RAND36) (age ≥ 18 years) [62].

    1. a.

      Measure for quality of life with 36 items.

  3. 3.

    Individualized Neuromuscular Quality of Life (INQoL) (age ≥ 18 years) [63].

    1. a.

      The INQoL is a validated muscle disease specific measure of quality of life, which can be used for individuals or large samples.

  4. 4.

    McGill pain questionnaire (age ≥ 12 years) [64].

    1. a.

      Questionnaire in which the location, level and characteristics of pain are assessed.

  5. 5.

    Wong-Baker Faces Pain rating scale (age ≥ 2 years) [65].

    1. a.

      The Wong-Baker Faces Pain Scale was originally created for children to help them communicate about their pain.

  6. 6.

    Checklist Individual Strength (CIS) (age ≥ 18 years) [66, 67].

    1. a.

      The CIS is a questionnaire rating four subscales: subjective tiredness, concentration, motivation and physical activity. It consists of 20 items on a seven-point scale.

  7. 7.

    ACTIVLIM (age ≥ 6 years) [68].

    1. a.

      Questionnaire to assess the ability to perform 22 activities of daily life on a three-point scale from impossible to easy.

  8. 8.

    Impact on Participation and Autonomy (IPA) (age ≥ 18 years) [69].

    1. a.

      Questionnaire about participation and autonomy in daily life.

  9. 9.

    Egen Klassifikation version 2 (EK2) (age ≥ 12 years and sufficient understanding of the English language) [70].

    1. a.

      The EK2 is a questionnaire that was designed to measure functional ability of activities in daily living in non-ambulant Duchenne muscular dystrophy patients. This questionnaire is available in English. Therefore, only patients who have a sufficient understanding of the English language will be asked to complete this questionnaire.

  10. 10.

    Borg Rating Scale of Perceived Exertion (Borg RPE scale) (age ≥ 5 years) [71].

    1. a.

      The Borg RPE scale is used to assess physical activity intensity level. In the LAST STRONG study participants are asked to assess their psychical sensations prior to and after the 6MWT.


At all four visits, muscle thickness and muscle echogenicity (quantitative grayscale analysis and Heckmatt ultrasound score) of a subset of bilateral muscles (See Table 2) will be assessed by muscle ultrasound using an Esaote MyLabTwice ultrasound scanner (Esaote SpA, Genoa, Italy) with an 3–13 MHz broadband linear transducer and a 53-mm footprint, adhering to a strictly defined and fixed measurement protocol [72,73,74,75,76,77,78]. To measure muscle echogenicity, the mean grayscale level within a manually selected region of interest (ROI) in the ultrasound image is calculated using an in-house developed software package in MATLAB (version 2013b, Mathworks, Natick, MA, USA). The muscle echogenicity and thickness are standardized by calculating their z-score: [[measured grayscale level – predicted grayscale level] / standard deviation grayscale]. The predicted grayscale level is calculated using a reference equation including age, length, sex and weight [78, 79]. In addition, all ultrasound images will be visually evaluated using the semi-quantitative Heckmatt grading scale [80]. At visit 1 (t = 0) and visit 3 (t = 12 months), muscle features are additionally qualitatively and (semi-)quantitatively described through full body muscle MRI (1,5 Tesla, Siemens, Erlangen, Germany) in accordance with our locally developed scanning protocol including Dixon vibe and Short-TI Inversion Recovery (STIR) images [81,82,83,84]. In accordance with previous studies on quantitative assessment of muscle MRI, the water and fat image of the Dixon sequence will be used to create a fat fraction map using MATLAB according to the following equation: Fat/(Fat + Water) [84]. The fat fraction map will be used to draw ROI (region of interest) per muscle using ImageJ software (ImageJ 1.47v, National Institutes of Health, USA). ROI will be drawn at predefined localization using the localizer sequences. All drawn ROI will be checked by a second clinician. Muscle cross-sectional area and fat fractions will be calculated per ROI. The qualitative and semi-quantitative assessment will be performed by two independent assessors. It will include assessment of a predefined subset of bilateral muscles (See Table 3), including relative reduction of muscle volume (0: no reduction, 1: mild, reduction of < 30%, 2: moderate, reduction of 30–60%, 3: severe, reduction of > 60%, 4: non-identifiable muscle), fatty infiltration (Modified Mercuri scale [85,86,87]), inflammation (absent/present, plus pattern: patchy, diffuse, peripheral/perifascial, central, other), and fibrosis (absent/present, plus pattern: fascial, intramuscular or other). A MRI will only be performed in patients of 10 years or older who are able to lie still for 45 min without respiratory equipment.

Table 2 Muscle ultrasound in a large subset of bilateral skeletal muscles
Table 3 Qualitative and semi-quantitative assessment of a large subset of bilateral skeletal muscles through MRI

At visit 1 (t = 0) and visit 3 (t = 12 months), an X-ray will be performed to assess deformities of the spine, including Cobb’s angle (spinal curvature), pelvic obliquity, coronal and sagittal balance and lumbar flexion and extension [88, 89]. In order to assess bone density, a DEXA-scan of the right femoral neck and lumbar spine, including a vertebral fracture assessment, will be performed at visit 1 (t = 0) and visit 3 (t = 12 months) by using the Hologic Discovery A Horizon DXA System (S/N 303053 M).

Cardiac assessment

In order to describe the prevalence and progression of cardiac comorbidities and the need for routine cardiac assessment, patients (age ≥ 2 years) will undergo electrocardiogram and conventional transthoracic echocardiography (TTE) with speckle tracking and Tissue Doppler Imaging (TDI) at visit 1 (t = 0) and visit 3 (t = 12 months). All investigations will be performed by EACVI TTE certified sonographers using commercially available ultrasound systems (Affiniti70 General, Philips Healthcare, Best, the Netherlands for adult participants; or Vivid E9 or Vivid E95, GE Healthcare Ultrasound, Horten, Norway for pediatric participants). Offline analysis will be performed using dedicated software (AGFA Enterprise Imaging Cardiology version 8.1.2, AGFA HealthCare, Mortsel, Belgium). Global Longitudinal strain (GLS) will be measured using speckle tracking echocardiography on a three beats acquisition with a frame rate > 60 frames/sec. All measurements will be done according to the EACVI recommendations for cardiac chamber quantification [90].

Pulmonary function

At all visits, patients (age ≥ 5 years) will undergo spirometry (forced expiratory volume in the first second (FEV1), forced vital capacity (FVC), vital capacity (VC), peak cough flow (PCF)) in upright and supine position (SpiroUSB, Vyaire Medical connected to PC Spirometry software, Spida CareFusion for Windows 7). Additionally, patients will undergo Maximum Expiratory Pressure (MEP), Maximum Inspiratory Pressure (MIP) and Sniff Nasal Inspiratory Pressure (SNIP) assessment in upright position (Micro RPM, Micro Medical, CareFusion, United Kingdom) [91]. Diaphragm ultrasound (MyLabTwice, Esaote SpA, Genoa, Italy) will be performed to assess diaphragm thickness (end expiratory and maximum inspiratory thickness) and thickening, and diaphragm echogenicity [92,93,94].


After all visits, patients (age ≥ 2 years) are asked to wear an accelerometer (GENEActiv Original, Activinsights Ltd) for eight consecutive days. In the same time period, patients or their parents are asked to fill in a diary with their major activities, including sleeping hours, physical exercise, work and school. The GENEActiv is a tri-axial, wrist-worn accelerometer that will be set to measure at 87,5 Hz sampling [95,96,97]. The raw data will be converted into 1-s epochs by using the GENEActiv Software (v.3.3, 2019). We will use the gravity subtracted sum of vector magnitudes (SVMgs) as the activity measure. The SVMgs will be measured using the following equation: SVMgs = ∑√(x2 + y2 + z2) -1 g. All data will be subsequently analyzed using MATLAB R2018a Update 4 ( for windows. A large subset of parameters will be addressed, including total activity (counts/day), the percentage of sedentary, light, moderate or vigorous activity, and total activity during sleep.

Statistical methods

Due to the explorative character of our study, we will use descriptive statistics (mean, median, SD, 95%-CI) in order to summarize our data. Further, Spearman’s correlation analysis and non-parametrical testing will be used to test the correlation between key parameters (i.e. age, ancillary investigations). Further, parameters will be corrected for genetic differences and partial or complete laminin subunit α2 deficiency. If reference values are available from literature, we will check for overlapping confidence intervals. In order to assess disease progression between subsequent measuring moments, we will perform the Wilcoxon signed-rank test (nonparametric continuous paired data) and the McNemar’s test (categorial paired data). Multiple linear regressions will be used to explore the relationship between potential disease modifying variables and disease severity. Linear mixed models will be applied for analysis of differences in disease progression. Further, we will correct for multiple testing. We regard p < 0,05 as statistically significant. The Statistical Package for the Social Sciences (SPSS version 25, IBM, Armonk, New York) will be used to conduct all statistical analyses.

Data collection

All data-management and data-monitoring will be performed within the Castor software (Version 2021) through direct entry or indirect entry via our electronic patient system (Epic, version May 2020).

Selection of outcome measures for future clinical trials

The results of our natural history study on both SELENON-RM and LAMA2-MD will feed an international key-opinion-leader workshop in which the in- and exclusion criteria, follow-up frequency and outcome measures for the international natural history studies will be discussed. When the results of preclinical and clinical work on promising new therapies continue to be encouraging, a clinical trial will be initiated.

Time plan

Patients will be included between August 2020 and August 2021 and will be followed for 1,5 year. The last visit of the last included patient is expected to take place in the beginning of 2023.


Here we present the design of the LAST STRONG study, an extensive natural history study in an unselected cohort of Dutch-speaking SELENON-RM and LAMA2-MD patients (both children and adults) that includes a broad plethora of clinical and functional outcome measures. This enables us to assess the clinical spectrum of patients diagnosed with SELENON-RM or LAMA2-MD. Hereby, we aim to fulfill our primary objectives namely, 1. to assess 1.5-year natural history in patients with SELENON-RM or LAMA2-MD; and 2. to select relevant and sensitive clinical and functional outcome measures to reach trial readiness. In addition, our insights will be vital for reaching our secondary objectives, including making a prevalence estimation, establishing a well-characterized baseline cohort and providing adequate symptomatic management leading to improved clinical care. We propose elaborate qualitative and quantitative measurements adapted to the age and functional abilities of the participant. The structured approach enables us to give an explorative, full-spectrum clinical description of patients diagnosed with SELENON-RM or LAMA2-MD. Further, there is a well-organized health system with full access to (pediatric) neurology and rehabilitation for every patient. Our center also has a longstanding history of patient recruitment for scientific research. Altogether, this is expected to reduce selection bias. Further, as a tertiary referral center for neuromuscular diseases, we are widely experienced with the clinical care and the conduction of studies in patients with rare neuromuscular diseases. Finally, all participants can reach our study center within a three-hour travel by car, which enables us to execute a population based, nationwide study. A consequence of our natural history study might be that the most severely affected patients may be less likely to participate in our study. We expect to overcome this problem to some extent by requesting all available medical records, sending out questionnaires and performing a medical history by video call/telephone or home visits for all patients that are not able or do not wish to visit our center, provided that written informed consent is given. Due to nationwide travel restrictions and local hospital measures related to the COVID-19 pandemic, Belgian patients are limited in their participation in our study.

The rarity of SELENON-RM and LAMA2-MD will result in a relatively small sample size of Dutch-speaking patients, which limits our possibilities to verify correlations or differences in subgroups. Consequently, we aim to contribute to international collaborations in order to enlarge the availability of natural history data.

The importance to reach trial readiness is warranted by the recent development of promising new treatment strategies for SELENON-RM and LAMA2-MD. Outcome measures need to be reliable and specified per age and disease severity for an adequate measurement of disease progression. If these outcome measures are not reliable, possible positive effects of future treatments in clinical trials will remain to be unrevealed. Reliable outcome measures are thus required in order to start clinical trials.

Further, deep phenotyping of our cohort provides us with valuable information on disease characteristics, enabling us to fulfil one of our secondary objectives, i.e. improving clinical care. For example, participants in whom clinically significant (co)morbidities are found during our study, are referred to the appropriate medical specialists. Moreover, we can take these (co)morbidities into account for routine clinical care of patients with SELENON-RM or LAMA2-MD that are not participating in our natural history study.

In our study, we do not take into account the selection of serum or urine biomarkers in order to limit the burden for patients participating in this study. However, in a study by Bharucha-Goebel et al. serum samples of patients with LAMA2-MD were analyzed for biomarkers. Proteins that were found to be altered, primarily consisted of cytokines and proteins involved in development and cell adhesion. Of the proteins decreased, most were cytokines, growth factors, and protease inhibitors [98]. We recommend to take serum or urine biomarkers into account in future studies.


The LAST STRONG study is expected to provide natural history data, which can be used for the selection of relevant clinical and functional outcome measures in order to reach trial readiness in SELONON-RM and LAMA2-MD patients. Further, our study aims to optimize clinical management in patients diagnosed with SELENON-RM or LAMA2-MD.

Availability of data and materials

The datasets generated during the current study will be available in the Donders Repository (


Borg RPE:

Borg Rating Scale of Perceived Exertion


The Children’s Hospital of Philadelphia Infant Test of Neuromuscular Disorders


Checklist Individual Strength


Dual Energy X-ray absorptiometry


Egen Klassifikation version 2


Functional Ambulation Classification


Forced expiratory volume in the first second


Forced vital capacity


Hammersmith functional motor scale


Hand-Held dynamometry


Hammersmith neurological examinations


Individualized Neuromuscular Quality of Life


Impact on Participation and Autonomy


LAMA2-related muscular dystrophy


Merosin-deficient congenital muscular dystrophy type 1A


Maximum Expiratory Pressure


Motor function measure – 20/32


Mini Balance Evaluation Systems Test


Maximum Inspiratory Pressure


Medical Research Council


Magnetic resonance imaging


Pediatric Balance Scale


Peak cough flow


Pediatric Quality of Life Inventory


Passive range of motion


Reasearch and Development-36


Region of interest


SELENON-related myopathy


Sniff Nasal Inspiratory Pressure


Short-TI Inversion Recovery


The gravity subtracted sum of vector magnitudes


Tissue doppler imaging


Transthoracic echocardiography


Vital capacity


6-Minute Walk Test


10-Meter Walk Test


  1. Witting N, Werlauff U, Duno M, Vissing J. Phenotypes, genotypes, and prevalence of congenital myopathies older than 5 years in Denmark. Neurol Genet. 2017;3(2):e140.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Villar-Quiles RN, von der Hagen M, Métay C, Gonzalez V, Donkervoort S, Bertini E, et al. The clinical, histologic, and genotypic spectrum of. Neurology. 2020;95(11):e1512–e27.

    Article  CAS  PubMed  Google Scholar 

  3. Nguyen Q, Lim KRQ, Yokota T. Current understanding and treatment of cardiac and skeletal muscle pathology in laminin-α2 chain-deficient congenital muscular dystrophy. Appl Clin Genet. 2019;12:113–30.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Geranmayeh F, Clement E, Feng LH, Sewry C, Pagan J, Mein R, et al. Genotype-phenotype correlation in a large population of muscular dystrophy patients with LAMA2 mutations. Neuromuscul Disord. 2010;20(4):241–50.

    Article  PubMed  Google Scholar 

  5. Sarkozy A, Foley AR, Zambon AA, Bönnemann CG, Muntoni F. LAMA2-related dystrophies: clinical phenotypes, disease biomarkers, and clinical trial readiness. Front Mol Neurosci. 2020;13:123.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Moghadaszadeh B, Petit N, Jaillard C, Brockington M, Quijano Roy S, Merlini L, et al. Mutations in SEPN1 cause congenital muscular dystrophy with spinal rigidity and restrictive respiratory syndrome. Nat Genet. 2001;29(1):17–8.

    Article  CAS  PubMed  Google Scholar 

  7. Ferreiro A, Quijano-Roy S, Pichereau C, Moghadaszadeh B, Goemans N, Bönnemann C, et al. Mutations of the selenoprotein N gene, which is implicated in rigid spine muscular dystrophy, cause the classical phenotype of multiminicore disease: reassessing the nosology of early-onset myopathies. Am J Hum Genet. 2002;71(4):739–49.

    Article  PubMed  PubMed Central  Google Scholar 

  8. Clarke NF, Kidson W, Quijano-Roy S, Estournet B, Ferreiro A, Guicheney P, et al. SEPN1: associated with congenital fiber-type disproportion and insulin resistance. Ann Neurol. 2006;59(3):546–52.

    Article  CAS  PubMed  Google Scholar 

  9. Ferreiro A, Ceuterick-de Groote C, Marks JJ, Goemans N, Schreiber G, Hanefeld F, et al. Desmin-related myopathy with Mallory body-like inclusions is caused by mutations of the selenoprotein N gene. Ann Neurol. 2004;55(5):676–86.

    Article  CAS  PubMed  Google Scholar 

  10. Wang CH, Bonnemann CG, Rutkowski A, Sejersen T, Bellini J, Battista V, et al. Consensus statement on standard of care for congenital muscular dystrophies. J Child Neurol. 2010;25(12):1559–81.

    Article  PubMed  PubMed Central  Google Scholar 

  11. Smeets HJM, Verbrugge B, Springuel P, Voermans NC, group MAW. International Workshop Report Congenital muscular dystrophy 1A: the road to therapy. Neuromuscul Disord. 2021.

  12. Janssen MCH, Koene S, de Laat P, Hemelaar P, Pickkers P, Spaans E, et al. The KHENERGY study: safety and efficacy of KH176 in mitochondrial m.3243A>G Spectrum disorders. Clin Pharmacol Ther. 2019;105(1):101–11.

    Article  CAS  PubMed  Google Scholar 

  13. Moulin M, Ferreiro A. Muscle redox disturbances and oxidative stress as pathomechanisms and therapeutic targets in early-onset myopathies. Semin Cell Dev Biol. 2017;64:213–23.

    Article  CAS  PubMed  Google Scholar 

  14. Arbogast S, Beuvin M, Fraysse B, Zhou H, Muntoni F, Ferreiro A. Oxidative stress in SEPN1-related myopathy: from pathophysiology to treatment. Ann Neurol. 2009;65(6):677–86.

    Article  CAS  PubMed  Google Scholar 

  15. Arbogast S, Ferreiro A. Selenoproteins and protection against oxidative stress: selenoprotein N as a novel player at the crossroads of redox signaling and calcium homeostasis. Antioxid Redox Signal. 2010;12(7):893–904.

    Article  CAS  PubMed  Google Scholar 

  16. Filipe A, Chernorudskiy A, Arbogast S, Varone E, Villar-Quiles RN, Pozzer D, et al. Defective endoplasmic reticulum-mitochondria contacts and bioenergetics in SEPN1-related myopathy. Cell Death Differ. 2021;28(1):123–38.

    Article  CAS  PubMed  Google Scholar 

  17. Pozzer D, Varone E, Chernorudskiy A, Schiarea S, Missiroli S, Giorgi C, et al. A maladaptive ER stress response triggers dysfunction in highly active muscles of mice with SELENON loss. Redox Biol. 2019;20:354–66.

    Article  CAS  PubMed  Google Scholar 

  18. Kemaladewi DU, Bassi PS, Erwood S, Al-Basha D, Gawlik KI, Lindsay K, et al. A mutation-independent approach for muscular dystrophy via upregulation of a modifier gene. Nature. 2019;572(7767):125–30.

    Article  CAS  PubMed  Google Scholar 

  19. Reinhard JR, Lin S, McKee KK, Meinen S, Crosson SC, Sury M, et al. Linker proteins restore basement membrane and correct LAMA2-related muscular dystrophy in mice. Sci Transl Med. 2017;9(396).

  20. Rooney JE, Knapp JR, Hodges BL, Wuebbles RD, Burkin DJ. Laminin-111 protein therapy reduces muscle pathology and improves viability of a mouse model of merosin-deficient congenital muscular dystrophy. Am J Pathol. 2012;180(4):1593–602.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Barraza-Flores P, Bates CR, Oliveira-Santos A, Burkin DJ. Laminin and integrin in LAMA2-related congenital muscular dystrophy: from disease to therapeutics. Front Mol Neurosci. 2020;13:1.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Chernorudskiy A, Varone E, Colombo SF, Fumagalli S, Cagnotto A, Cattaneo A, et al. Selenoprotein N is an endoplasmic reticulum calcium sensor that links luminal calcium levels to a redox activity. Proc Natl Acad Sci U S A. 2020;117(35):21288–98.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Patton BL, Miner JH, Chiu AY, Sanes JR. Distribution and function of laminins in the neuromuscular system of developing, adult, and mutant mice. J Cell Biol. 1997;139(6):1507–21.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Petrof BJ, Shrager JB, Stedman HH, Kelly AM, Sweeney HL. Dystrophin protects the sarcolemma from stresses developed during muscle contraction. Proc Natl Acad Sci U S A. 1993;90(8):3710–4.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Fontes-Oliveira CC, Steinz M, Schneiderat P, Mulder H, Durbeej M. Bioenergetic impairment in congenital muscular dystrophy type 1A and Leigh syndrome muscle cells. Sci Rep. 2017;7(1):45272.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Harandi VM, Oliveira BMS, Allamand V, Friberg A, Fontes-Oliveira CC, Durbeej M. Antioxidants Reduce Muscular Dystrophy. Antioxidants. 2020;9(3).

  27. Kemaladewi D, Hyatt E, Ivakine Z, Cohn R. CRISPR/Cas9-mediated exon inclusion in Lama2 gene alleviates dystrophic pathology in MDC1A mouse model in title abstract keyword. Neuromuscul Disord. 2016;26:S190.

    Article  Google Scholar 

  28. Wright M. Calcium handling in a zebrafish model of SELENON congenital muscular dystrophy: World Muscle Society: Neuromuscular Disorders; 2020. p. S149–S50.

  29. Deniziak M, Thisse C, Rederstorff M, Hindelang C, Thisse B, Lescure A. Loss of selenoprotein N function causes disruption of muscle architecture in the zebrafish embryo. Exp Cell Res. 2007;313(1):156–67.

    Article  CAS  PubMed  Google Scholar 

  30. Bachmann C, Noreen F, Voermans NC, Schär PL, Vissing J, Fock JM, et al. Aberrant regulation of epigenetic modifiers contributes to the pathogenesis in patients with selenoprotein N-related myopathies. Hum Mutat. 2019;40(7):962–74.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Marino M, Stoilova T, Giorgi C, Bachi A, Cattaneo A, Auricchio A, et al. SEPN1, an endoplasmic reticulum-localized selenoprotein linked to skeletal muscle pathology, counteracts hyperoxidation by means of redox-regulating SERCA2 pump activity. Hum Mol Genet. 2015;24(7):1843–55.

    Article  CAS  PubMed  Google Scholar 

  32. Yurchenco PD, McKee KK, Reinhard JR, Rüegg MA. Laminin-deficient muscular dystrophy: Molecular pathogenesis and structural repair strategies. Matrix Biol. 2018;71–72:174–87.

    Article  Google Scholar 

  33. Hara Y, Mizobe Y, Miyatake S, Takizawa H, Nagata T, Yokota T, et al. Exon skipping using antisense oligonucleotides for laminin-Alpha2-deficient muscular dystrophy. Methods Mol Biol. 1828;2018:553–64.

    Google Scholar 

  34. Hall TE, Wood AJ, Ehrlich O, Li M, Sonntag CS, Cole NJ, et al. Cellular rescue in a zebrafish model of congenital muscular dystrophy type 1A. NPJ Regen Med. 2019;4(1):21.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Meilleur KG, Jain MS, Hynan LS, Shieh CY, Kim E, Waite M, et al. Results of a two-year pilot study of clinical outcome measures in collagen VI- and laminin alpha2-related congenital muscular dystrophies. Neuromuscul Disord. 2015;25(1):43–54.

    Article  PubMed  Google Scholar 

  36. Jain MS, Meilleur K, Kim E, Norato G, Waite M, Nelson L, et al. Longitudinal changes in clinical outcome measures in COL6-related dystrophies and LAMA2-related dystrophies. Neurology. 2019;93(21):e1932–e43.

    Article  PubMed  PubMed Central  Google Scholar 

  37. Zambon AA, Ridout D, Main M, Mein R, Phadke R, Muntoni F, et al. LAMA2-related muscular dystrophy: natural history of a large pediatric cohort. Ann Clin Transl Neurol. 2020;7(10):1870–82.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Beenakker EA, van der Hoeven JH, Fock JM, Maurits NM. Reference values of maximum isometric muscle force obtained in 270 children aged 4-16 years by hand-held dynamometry. Neuromuscul Disord. 2001;11(5):441–6.

    Article  CAS  PubMed  Google Scholar 

  39. van den Beld WA, van der Sanden GA, Sengers RC, Verbeek AL, Gabreëls FJ. Validity and reproducibility of hand-held dynamometry in children aged 4-11 years. J Rehabil Med. 2006;38(1):57–64.

    Article  PubMed  Google Scholar 

  40. McKay MJ, Baldwin JN, Ferreira P, Simic M, Vanicek N, Burns J, et al. Normative reference values for strength and flexibility of 1,000 children and adults. Neurology. 2017;88(1):36–43.

    Article  PubMed  PubMed Central  Google Scholar 

  41. van der Ploeg RJ, Fidler V, Oosterhuis HJ. Hand-held myometry: reference values. J Neurol Neurosurg Psychiatry. 1991;54(3):244–7.

    Article  PubMed  PubMed Central  Google Scholar 

  42. Soucie JM, Wang C, Forsyth A, Funk S, Denny M, Roach KE, et al. Range of motion measurements: reference values and a database for comparison studies. Haemophilia. 2011;17(3):500–7.

    Article  CAS  PubMed  Google Scholar 

  43. Glanzman AM, Mazzone E, Main M, Pelliccioni M, Wood J, Swoboda KJ, et al. The Children's Hospital of Philadelphia infant test of neuromuscular disorders (CHOP INTEND): test development and reliability. Neuromuscul Disord. 2010;20(3):155–61.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Glanzman AM, McDermott MP, Montes J, Martens WB, Flickinger J, Riley S, et al. Validation of the Children’s Hospital of Philadelphia infant test of neuromuscular disorders (CHOP INTEND). Pediatr Phys Ther. 2011;23(4):322–6.

    Article  Google Scholar 

  45. Dubowitz L, Ricciw D, Mercuri E. The Dubowitz neurological examination of the full-term newborn. Ment Retard Dev Disabil Res Rev. 2005;11(1):52–60.

    Article  PubMed  Google Scholar 

  46. de Lattre C, Payan C, Vuillerot C, Rippert P, de Castro D, Bérard C, et al. Motor function measure: validation of a short form for young children with neuromuscular diseases. Arch Phys Med Rehabil. 2013;94(11):2218–26.

    Article  PubMed  Google Scholar 

  47. Bérard C, Payan C, Hodgkinson I, Fermanian J, Group MCS. A motor function measure for neuromuscular diseases. Construction and validation study. Neuromuscul Disord. 2005;15(7):463–70.

    Article  PubMed  Google Scholar 

  48. Main M, Kairon H, Mercuri E, Muntoni F. The Hammersmith functional motor scale for children with spinal muscular atrophy: a scale to test ability and monitor progress in children with limited ambulation. Eur J Paediatr Neurol. 2003;7(4):155–9.

    Article  PubMed  Google Scholar 

  49. Ramsey D, Scoto M, Mayhew A, Main M, Mazzone ES, Montes J, et al. Revised Hammersmith scale for spinal muscular atrophy: a SMA specific clinical outcome assessment tool. PLoS One. 2017;12(2):e0172346.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Franjoine MR, Gunther JS, Taylor MJ. Pediatric balance scale: a modified version of the berg balance scale for the school-age child with mild to moderate motor impairment. Pediatr Phys Ther. 2003;15(2):114–28.

    Article  PubMed  Google Scholar 

  51. Franchignoni F, Horak F, Godi M, Nardone A, Giordano A. Using psychometric techniques to improve the balance evaluation systems test: the mini-BESTest. J Rehabil Med. 2010;42(4):323–31.

    Article  PubMed  PubMed Central  Google Scholar 

  52. Florence JM, Pandya S, King WM, Robison JD, Signore LC, Wentzell M, et al. Clinical trials in Duchenne dystrophy. Standardization and reliability of evaluation procedures. Phys Ther. 1984;64(1):41–5.

    Article  CAS  PubMed  Google Scholar 

  53. Mayhew JE, Florence JM, Mayhew TP, Henricson EK, Leshner RT, McCarter RJ, et al. Reliable surrogate outcome measures in multicenter clinical trials of Duchenne muscular dystrophy. Muscle Nerve. 2007;35(1):36–42.

    Article  PubMed  Google Scholar 

  54. Enright PL. The six-minute walk test. Respir Care. 2003;48(8):783–5.

    Google Scholar 

  55. Holden MK, Gill KM, Magliozzi MR, Nathan J, Piehl-Baker L. Clinical gait assessment in the neurologically impaired. Reliability and meaningfulness. Phys Ther. 1984;64(1):35–40.

    Article  CAS  PubMed  Google Scholar 

  56. Jung IY, Chae JH, Park SK, Kim JH, Kim JY, Kim SJ, et al. The correlation analysis of functional factors and age with duchenne muscular dystrophy. Ann Rehabil Med. 2012;36(1):22–32.

    Article  PubMed  PubMed Central  Google Scholar 

  57. Brooke MH, Griggs RC, Mendell JR, Fenichel GM, Shumate JB, Pellegrino RJ. Clinical trial in Duchenne dystrophy. I. the design of the protocol. Muscle Nerve. 1981;4(3):186–97.

    Article  CAS  PubMed  Google Scholar 

  58. Lue YJ, Lin RF, Chen SS, Lu YM. Measurement of the functional status of patients with different types of muscular dystrophy. Kaohsiung J Med Sci. 2009;25(6):325–33.

    Article  PubMed  Google Scholar 

  59. Engelen V, Haentjens MM, Detmar SB, Koopman HM, Grootenhuis MA. Health related quality of life of Dutch children: psychometric properties of the PedsQL in the Netherlands. BMC Pediatr. 2009;9(1):68.

    Article  PubMed  PubMed Central  Google Scholar 

  60. Iannaccone ST, Hynan LS, Morton A, Buchanan R, Limbers CA, Varni JW, et al. The PedsQL in pediatric patients with spinal muscular atrophy: feasibility, reliability, and validity of the pediatric quality of life inventory generic Core scales and neuromuscular module. Neuromuscul Disord. 2009;19(12):805–12.

    Article  PubMed  PubMed Central  Google Scholar 

  61. Gordijn M, Suzanne Gordijn M, Cremers EM, Kaspers GJ, Gemke RJ. Fatigue in children: reliability and validity of the Dutch PedsQL™ multidimensional fatigue scale. Qual Life Res. 2011;20(7):1103–8.

    Article  PubMed  Google Scholar 

  62. Aaronson NK, Muller M, Cohen PD, Essink-Bot ML, Fekkes M, Sanderman R, et al. Translation, validation, and norming of the Dutch language version of the SF-36 health survey in community and chronic disease populations. J Clin Epidemiol. 1998;51(11):1055–68.

    Article  CAS  PubMed  Google Scholar 

  63. Seesing FM, van Vught LE, Rose MR, Drost G, van Engelen BG, van der Wilt GJ. The individualized neuromuscular quality of life questionnaire: cultural translation and psychometric validation for the Dutch population. Muscle Nerve. 2015;51(4):496–500.

    Article  PubMed  Google Scholar 

  64. Vanderiet K, Adriaensen H, Carton H, Vertommen H. The McGill pain questionnaire constructed for the Dutch language (MPQ-DV). Preliminary data concerning reliability and validity. Pain. 1987;30(3):395–408.

    Article  PubMed  Google Scholar 

  65. Wong DL, Baker CM. Pain in children: comparison of assessment scales. Pediatr Nurs. 1988;14(1):9–17.

    CAS  PubMed  Google Scholar 

  66. Worm-Smeitink M, Gielissen M, Bloot L, van Laarhoven HWM, van Engelen BGM, van Riel P, et al. The assessment of fatigue: psychometric qualities and norms for the checklist individual strength. J Psychosom Res. 2017;98:40–6.

    Article  CAS  PubMed  Google Scholar 

  67. Vercoulen JH, Swanink CM, Fennis JF, Galama JM, van der Meer JW, Bleijenberg G. Dimensional assessment of chronic fatigue syndrome. J Psychosom Res. 1994;38(5):383–92.

    Article  CAS  PubMed  Google Scholar 

  68. Vandervelde L, Van den Bergh PY, Goemans N, Thonnard JL. ACTIVLIM: a Rasch-built measure of activity limitations in children and adults with neuromuscular disorders. Neuromuscul Disord. 2007;17(6):459–69.

    Article  PubMed  Google Scholar 

  69. Cardol M, de Haan RJ, van den Bos GA, de Jong BA, de Groot IJ. The development of a handicap assessment questionnaire: the impact on participation and autonomy (IPA). Clin Rehabil. 1999;13(5):411–9.

    Article  CAS  PubMed  Google Scholar 

  70. Steffensen B, Hyde S, Lyager S, Mattsson E. Validity of the EK scale: a functional assessment of non-ambulatory individuals with Duchenne muscular dystrophy or spinal muscular atrophy. Physiother Res Int. 2001;6(3):119–34.

    Article  CAS  PubMed  Google Scholar 

  71. Borg GA. Psychophysical bases of perceived exertion. Med Sci Sports Exerc. 1982;14(5):377–81.

    Article  CAS  Google Scholar 

  72. van Alfen N, Mah JK. Neuromuscular ultrasound: a new tool in your toolbox. Can J Neurol Sci. 2018;45(5):504–15.

    Article  PubMed  Google Scholar 

  73. Mah JK, van Alfen N. Neuromuscular ultrasound: clinical applications and diagnostic values. Can J Neurol Sci. 2018;45(6):605–19.

    Article  PubMed  Google Scholar 

  74. Pillen S, van Alfen N. Skeletal muscle ultrasound. Neurol Res. 2011;33(10):1016–24.

    Article  PubMed  Google Scholar 

  75. Pillen S, Van Alfen N. Muscle ultrasound from diagnostic tool to outcome measure--quantification is the challenge. Muscle Nerve. 2015;52(3):319–20.

    Article  PubMed  Google Scholar 

  76. Pillen S, Verrips A, van Alfen N, Arts IM, Sie LT, Zwarts MJ. Quantitative skeletal muscle ultrasound: diagnostic value in childhood neuromuscular disease. Neuromuscul Disord. 2007;17(7):509–16.

    Article  CAS  PubMed  Google Scholar 

  77. Pillen S, Boon A, Van Alfen N. Muscle ultrasound. Handb Clin Neurol. 2016;136:843–53.

    Article  PubMed  Google Scholar 

  78. Wijntjes J, van Alfen N. Muscle ultrasound: present state and future opportunities. Muscle Nerve. 2021;63(4):455–66.

    Article  PubMed  Google Scholar 

  79. Scholten RR, Pillen S, Verrips A, Zwarts MJ. Quantitative ultrasonography of skeletal muscles in children: normal values. Muscle Nerve. 2003;27(6):693–8.

    Article  CAS  PubMed  Google Scholar 

  80. Heckmatt JZ, Leeman S, Dubowitz V. Ultrasound imaging in the diagnosis of muscle disease. J Pediatr. 1982;101(5):656–60.

    Article  CAS  PubMed  Google Scholar 

  81. Dahlqvist JR, Widholm P, Leinhard OD, Vissing J. MRI in neuromuscular diseases: an emerging diagnostic tool and biomarker for prognosis and efficacy. Ann Neurol. 2020;88(4):669–81.

    Article  PubMed  Google Scholar 

  82. Morrow JM, Sinclair CD, Fischmann A, Machado PM, Reilly MM, Yousry TA, et al. MRI biomarker assessment of neuromuscular disease progression: a prospective observational cohort study. Lancet Neurol. 2016;15(1):65–77.

    Article  PubMed  PubMed Central  Google Scholar 

  83. Mul K, Horlings CGC, Vincenten SCC, Voermans NC, van Engelen BGM, van Alfen N. Quantitative muscle MRI and ultrasound for facioscapulohumeral muscular dystrophy: complementary imaging biomarkers. J Neurol. 2018;265(11):2646–55.

    Article  PubMed  PubMed Central  Google Scholar 

  84. Mul K, Vincenten SCC, Voermans NC, Lemmers RJLF, van der Vliet PJ, van der Maarel SM, et al. Adding quantitative muscle MRI to the FSHD clinical trial toolbox. Neurology. 2017;89(20):2057–65.

    Article  PubMed  PubMed Central  Google Scholar 

  85. Kinali M, Arechavala-Gomeza V, Cirak S, Glover A, Guglieri M, Feng L, et al. Muscle histology vs MRI in Duchenne muscular dystrophy. Neurology. 2011;76(4):346–53.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  86. Mercuri E, Cini C, Pichiecchio A, Allsop J, Counsell S, Zolkipli Z, et al. Muscle magnetic resonance imaging in patients with congenital muscular dystrophy and Ullrich phenotype. Neuromuscul Disord. 2003;13(7–8):554–8.

    Article  CAS  PubMed  Google Scholar 

  87. Mercuri E, Pichiecchio A, Counsell S, Allsop J, Cini C, Jungbluth H, et al. A short protocol for muscle MRI in children with muscular dystrophies. Eur J Paediatr Neurol. 2002;6(6):305–7.

    Article  PubMed  Google Scholar 

  88. Gupta MC, Wijesekera S, Sossan A, Martin L, Vogel LC, Boakes JL, et al. Reliability of radiographic parameters in neuromuscular scoliosis. Spine (Phila Pa 1976). 2007;32(6):691–5.

    Article  Google Scholar 

  89. Kim H, Kim HS, Moon ES, Yoon CS, Chung TS, Song HT, et al. Scoliosis imaging: what radiologists should know. Radiographics. 2010;30(7):1823–42.

    Article  PubMed  Google Scholar 

  90. Lang RM, Badano LP, Mor-Avi V, Afilalo J, Armstrong A, Ernande L, et al. Recommendations for cardiac chamber quantification by echocardiography in adults: an update from the American Society of Echocardiography and the European Association of Cardiovascular Imaging. Eur Heart J Cardiovasc Imaging. 2015;16(3):233–70.

    Article  Google Scholar 

  91. Laveneziana P, Albuquerque A, Aliverti A, Babb T, Barreiro E, Dres M, et al. ERS statement on respiratory muscle testing at rest and during exercise. Eur Respir J. 2019;53(6).

  92. Fayssoil A, Nguyen LS, Ogna A, Stojkovic T, Meng P, Mompoint D, et al. Diaphragm sniff ultrasound: Normal values, relationship with sniff nasal pressure and accuracy for predicting respiratory involvement in patients with neuromuscular disorders. PLoS One. 2019;14(4):e0214288.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  93. Caggiano S, Khirani S, Dabaj I, Cavassa E, Amaddeo A, Arroyo JO, et al. Diaphragmatic dysfunction in SEPN1-related myopathy. Neuromuscul Disord. 2017;27(8):747–55.

    Article  PubMed  Google Scholar 

  94. van Doorn JLM, Pennati F, Hansen HHG, van Engelen BGM, Aliverti A, Doorduin J. Respiratory muscle imaging by ultrasound and MRI in neuromuscular disorders. Eur Respir J. 2021.

  95. Esliger DW, Rowlands AV, Hurst TL, Catt M, Murray P, Eston RG. Validation of the GENEA accelerometer. Med Sci Sports Exerc. 2011;43(6):1085–93.

    Article  PubMed  Google Scholar 

  96. Phillips LR, Parfitt G, Rowlands AV. Calibration of the GENEA accelerometer for assessment of physical activity intensity in children. J Sci Med Sport. 2013;16(2):124–8.

    Article  PubMed  Google Scholar 

  97. de Vries PR, Janssen M, Spaans E, de Groot I, Janssen A, Smeitink J, et al. Natural variability of daily physical activity measured by accelerometry in children with a mitochondrial disease. Mitochondrion. 2019;47:30–7.

    Article  CAS  PubMed  Google Scholar 

  98. Bharucha-Goebel D, Collins J, Hu Y, Foley AR, Donkervoort S, Leach M, et al. Serum biomarker discovery for congenital muscular dystrophies. Neuromuscul Disord. 2016.

Download references


We thank all patients and their relatives for participation in our study. Several authors of this publication are members of the Netherlands Neuromuscular Center (NL-NMD) and the European Reference Network for rare neuromuscular diseases (EURO-NMD).


The study is financially supported by a competitively awarded, peer-reviewed grant from Stichting Spieren voor Spieren, Stichting Stofwisselkracht and Stichting Voor Sara, The Netherlands. The scientific advisory boards of the aforementioned foundations took into account the scientific quality of the study protocol, impact of the study, quality of the applicants and requested support. The funders have no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The content is solely the responsibility of the authors and does not necessarily represent the official views of the Stichting Spieren voor Spieren, Stichting Stofwisselkracht and Stichting Voor Sara.

Author information

Authors and Affiliations



KB, JTG, JD, NA, FEAU, FMAH, RN, WCMT, SCFMB, ATMD, JMTD, MCHJ, EK, ESBK, SK, JAMS, BK, FHJT, HJMS, BGME, CEE and NCV contributed to the study conception and design. Further, KB, JTG, CEE, NCV drafted the manuscript or revised it critically. All authors have read and approved the final manuscript and agreed to be accountable for their contributions.

Corresponding author

Correspondence to Karlijn Bouman.

Ethics declarations

Ethics approval and consent to participate

This study is performed in accordance with the Declaration of Helsinki and has been approved by the medical ethical reviewing committee of Region Arnhem-Nijmegen (NL-number NL64269.091.17, dossier number 2017–3911; date of approval of last amendment: 8 July 2020). From all patients, or in case of children their parents or legal guardian, written informed consent will be obtained for participating in our study.

Consent for publication

Not applicable.

Competing interests

Professor Jan A.M. Smeitink is Chief Executive Officer of Khondrion. All other authors declare that they have no competing interests.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit The Creative Commons Public Domain Dedication waiver ( applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bouman, K., Groothuis, J.T., Doorduin, J. et al. Natural history, outcome measures and trial readiness in LAMA2-related muscular dystrophy and SELENON-related myopathy in children and adults: protocol of the LAST STRONG study. BMC Neurol 21, 313 (2021).

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: